An English Medium Co. Ed.

Senior

Secondary School

Investigatory
Project
On

SUBMITTED BY:
SUBMITTED TO:
Su#hag Singh Mr.
Sandee$ Kulshesthra
XII Sci. B
(H.O.D Biology)

A*no+ledgement
I am over+helmed in all hum#leness and gratefulness to ac*no+ledge
m y de$th to all those + h o have hel$ed m e to $ut these ideas- + ell
a#ove the level of sim$licity and into something concrete.
I + o u l d li*e to ex$ress m y s$ecial t han* s of gratitude to m y #iology
teacher- Mr. S a n d e e p K u l s h e s t h r a as +ell as our Princi$al M r s . N i d h i
B h a t i a +ho gave me the golden o$$ortunity to do this
+onderful
$roject on the to$ic /A$$lications of Biotechnology0- +hich also hel$ed
me in doing a lot of research and I came to * n o + a#out so many n e +
things. I am really than*ful to them.
Any a tt em$t at any level can1t #e satisfactorily com$leted + i t h o u t the
su$$ort and guidance of m y P a r e n t s a n d F r i e n d s + h o hel$ed me a lot
in gathering different information- collecting data and guiding me from
time to time in ma*ing this $roject- des$ite of their #usy schedules- they
gave me different ideas in ma*ing this $roject uni3ue. I am than*ful to
them too.
I am m a * i n g this $roject not only for m a r * s #ut to also increase m y
*no+ledge...
Than*ing you
Subhag Singh
XII Sci. B
 Certificate
This is to certify that SUBHAG SINGH of class
X I I SCI.B of GYAN DEEP SHIKSHA BHARATI
has successfully com$leted the
investigatory
$roject on the to$ic ”APPLICATIONS OF
BIOTECHNOLOGY” under the guidance
of
MR. SANDEEP KULSHESTHRA
(H.O.D.
Biology) during the session 5678-76 in
the
$artial fulfilment of Biology
Practical
Examination conducted #y CENTRAL
BOARD OF SECONDARY EDUCATION
(AISSCE).
Mr. Sandeep Kulshesthra External
Examiner
(H.O.D Biology) (C.B.S.E)

Intr
odu
ctio
B i o t e c h n o l o g y is the use of living systems and organisms to develop or
make products, or "any technological application that uses biological
systems, living organisms or derivatives thereof, to make or modify
products
n or processes for specific use .

At its simplest, biotechnology

is technology based on biology -
W h a t is biotechnology harnesses
Biotechnology? cellular and molecular
bio develop
processes to and
technologies that help
products
improve our lives and the
health of our planet.
We have used the
biological processes of
microorganisms for more than
6,000 years to make
useful food products, such as bread and cheese, and to preserve
dairy products.

Modern biotechnology provides breakthrough products and

technologies to combat debilitating and rare diseases, reduce our
environmental footprint, feed the hungry, useless and cleaner energy,
and have safer, cleaner and more efficient industrial manufacturing
processes.
 Biotech is hel$ing to heal the +orld #y harnessing nat ure?s o + n tool#ox
and using our o + n genetic ma*eu$ to heal and guide lines of research
#y:
• @educing rates of infectious disease
• Saving millions of children?s lives
• Changing the odds of serious- life-threatening conditions affecting
millions around the +orld
• Tailoring treatments to individuals to minimize health ris*s and
side effects
• Creating more $recise tools for disease detection
• Com#ating serious illnesses and everyday threats confronting the
develo$ing +orld.

History
Throughout the history of agriculture- farmers have inadvertently
altered the genetics of their cro$s through introducing th em to n e +
environments and #reeding t h e m + i t h other $lants - one of the first
forms of #iotechnology.
These processes also were included in early f e r m e n t a t i o n o f b e er .
In brewing, malted grains (containing enzymes) convert starch from
grains into sugar and then adding
specific yeasts to produce beer. In
this process, carbohydrates in the
grains were broken down into
alcohols such as ethanol. Later
other cultures produced the
process of lactic acid
fermentation which allowed the
fermentation and preservation of
other forms of food, such as s oy
sauce. Fermentation was also
used in this time period to produce I e a v e n e d b r e a d . Although the
process of fermentation was not fully understood until Louis Pasteur's
work in 1857, it is still the first use of biotechnology to convert a food
source into another form.

For thousands of years, humans have used selective breeding to

improve production of crops and livestock to use them for food. In
selective breeding, organisms with desirable characteristics are mated
to produce offspring with the same characteristics. For example, this
technique was used with corn to produce the largest and sweetest
crops.

Biotechnology has also led t o the development of antibiotics. In

1928, Alexander Fleming discovered the mould Pen iciI I ium . His work
led to the purification of the antibiotic compound formed by the mould
by Howard Florey, Ernst Boris Chain and Norman Heatley - to form what
we today know as penicillin. In 1940, penicillin became available for
medicinal use to treat bacterial infections in humans.

The field of modern biotechnology is generally thought of as having

been born in 1971 when Paul Berg's experiments in g e n e s p I i c i n g had
early success. Herbert W. Boyer and Stanley N. Cohen significantly
advanced the new technology in 1972 by transferring genetic material
into a bacterium, such that the imported material would be reproduced.
 Biotechnology in
Agriculture
Genetically M od if ie d Crops

G e n e t i c a l l y m o d i f i e d c r o p s or /GM
cro$s0 or /#iotech cro$s0 are $lants used
in agriculture- the DGA of +hich has #een
modified

+ i t h genetic
engineering techni3ues. In most cases
the aim is to introduce a n e + trait to the
$lant + h i c h does not occur naturally in
the s$ecies. Exam$les in food cro$s
include resistance to certain $ests- diseases- stressful environmental
conditions- resistance to chemical treatments- reduction of s$oilage- or
im$roving the nutrient $rofile of the cro$. Exam$les in non-food cro$s
include $roduction of $harmaceutical agents- #io fuels- and other
industrially useful goods- as +ell as for #ioremediat ion .

Plant s and cro$s + it h GM t rait s have #een t est ed m ore t han any ot her
cro$sJ + it h no credi#le evidence of harm t o hum ans or anim als. In fact -
seeds + it h GM t rait s have #een t est ed m ore t han any ot her cro$s in t he
hist ory of agricult ure K + it h no credi#le evidence of harm t o hum ans or
anim als.

Governmental regulatory agencies- scientific organizations and leading

health associations +orld+ide agree that food gro+n from GM c r o p s is
s a f e t o e a t . The = o r l d Health Organization- the American Medical
Association- the U.S. Gational Academy of Sciences- the British @oyal
Society- among others that have examined the evidence- all come to the
same conclusion: consuming foods containing ingredients derived from
GM cro$s is safe to eat and no ris*ier than consuming the same foods
containing ingredients from cro$ $lants modified #y conventional $lant
im$rovement techni3ues. Genetic modifications have:

7. Made cro$s more tolerant to a#iotic stresses (cold- drought- salt-

heat).

5. @educed reliance on chemical $esticides ($est resistant cro$s).

 L. Hel$ed to reduce $ost harvest losses  enhanced the nutritional
value of the food.

#NA Inter/erence )#NAi*

@GA interference (@GAi) is a method of #loc*ing gene function #y
inserting short se3uences of ri#onucleic acid (@GA) that match $art of
the target gene1s se3uence- thus no $roteins are $roduced. @GAi has the
$otential to #ecome a $o+erful thera$eutic a$$roach t o + a r d targeted
and $ersonalized medicine. @GAi has $rovided a + a y to control $ests
and diseases- introduce novel $lant traits and increase cro$ yield. Using
@GAi- scientists have develo$ed novel cro$s such as nicotine-free
to#acco- non-allergenic $eanuts- decaffeinated coffee- and nutrient
fortified maize among many others.

Mechanism of @GA interferences as understood is that it comes into $lay

+ h e n a dou#le stranded @GA is introduced either naturally or artificially
in a cell. An endo ri#onuclease enzyme cleaves the long ds@GA into
small $ieces of @GA. The small $ieces could #e m i @GA or si @GA
de$ending u$on the origin of long ds@GA i.e. endogenous or exogenous
res$ectively. A dou#le
stranded @GA may #e
generated #y either @GA
de$endent @GA $olymerase
or #idirectional
transcri$tion of trans$osa#le
elements or
$hysically introduced.
There are o$ several
$ortunities for the
a$$lications of in
@GAi science
cro$

for

its

im$rovement such as stress

tolerance and
enhanced nutritional level.This * n oc * d o + n technology may #e useful in
inducing early flo+ering- delayed ri$ening- delayed senescence-
#rea*ing dormancy- stress-free $lants- overcoming self-sterility- etc .
genome-+ i d e identification of gene function that should hel$ im$rove
our understanding of $lant $arasitic nematodes. @GAi should hel$
identify gene and- hence- $rotein targets for nematode control
strategies. Pros$ects for novel resistance de$end on the $lant
generating an effective form of dou#le-stranded @GA in the a#sence of
an endogenous target gene +ithout detriment to itself. These @GA
molecules must then #ecome availa#le to the nematode and #e ca$a#le
of ingestion via its feeding tu#e. If these re3uirements can #e met- cro$
resistance could #e achieved #y a $lant delivering a ds@GA that targets
a nematode gene and induces a lethal or highly damaging @GAi effect
on the $arasit e .

Bt to3in
A $rotein that is toxic to che+ing insects and is $roduced #y the soil
#acterium Bacillus thuringiensis and has long #een used as a #iological
p e s t i c i d e . By means of genetic engineering- the genes for Bt toxin can
#e isolated from Bacillus thuringiensis and transferred to $lants.

B a c i l l u s t h u r i n g i e n s i s ) B t * is a #acteria that $roduces $roteins +hich

are toxic to insects. But extreme toxicity comes at no sur$rise. It 1s in the
same family of #acteria as B. anthracis- + hic h causes anthrax- and B.
cereus- +hich causes food $oisoning.

The Bt toxin dissolve in the high $H insect gut and #ecome active. The
toxins then a t t a c * the gut cells of the insect- $unching holes in the
lining. The Bt s$ores s$ills out of the gut and germinate in the insect
causing death +ithin a cou$le days.

Even though the toxin does not *ill the insect immediately- treated $lant
$arts +ill not #e damaged #ecause the insect sto$s feeding +ithin
hours. Bt s$ores do not s$read to other insects or cause disease
out#rea*s on their o+n.
7. Insect eats Bt crystals and s$ores.

5. The toxin #inds to s$ecific

rece$tors in the gut and the insects
sto$s eating.

L. The crystals cause the gut + all

to #rea* do+n- a lo+ing s$ores
and normal gut #acteria to enter the
#ody.

H. The insect dies as s$ores and

gut #acteria $roliferate in the #ody.

Bt action is very s$ecific. Different strains of Bt are s$ecific to different

rece$tors in insect gut + a l. Bt toxicity de$ends on recognizing
rece$tors- damage to the gut #y the toxin occurs u$on #inding to a
rece$tor. Each insect s$ecies $ossesses different ty$es of rece$tors that
+ill match only certain toxin $roteins- li*e a loc* to a *ey.

It is #ecause of this that farmers have to #e careful to match the target

$est s$ecies + i t h a $articular Bt toxin $rotein +hich is s$ecific for that
insect. This also hel$s the #enifical insects #ecause they +ill usually not
#e harmed #y that $articular strain of Bt .

Bt Cotton
B t c o t t o n is a genetically modified organism (GMO) cotton variety,
which produces an insecticide to bollworm. Strains of the
bacterium Bacillus thuringiensis produce over 200 different Bt toxins,
each harmful to different insects. Most notably,
Bt toxins are insecticidal to the larvae of moths
and butterflies, beetles, cotton bollworms and
ghtu flies but are harmless to other forms of life.
The gene coding for Bt toxin has been inserted
into cotton as a transgene, causing it to produce
this natural insecticide in its tissues. In many
regions, the main pests in commercial cotton
are lepidopteran larvae, which are killed by the
Bt protein in thegenetically modified cotton they
eat. This eliminates the need t o use large
amounts of broad-spectrum insecticides to kill lepidopteran pests. This
spares natural insect predators in the farm ecology and further
contributes to non insecticide pest management.

Bt cotton is ineffective against many cotton pests such as plant

bugs, stink bugs, and aphids; depending on circumstances it ma y be
desirable t o use insecticides in prevention. A 2006 study done
by Cornell researchers, the Center for
Chinese Agricultural Policy and the Chinese
Academy of Science on Bt cotton farming in
China found that after seven years these
secondary pests that were normally
controlled by pesticide had increased,
necessitating the use of pesticides at similar
levels to non-Bt cotton and causing less
profit for farmers because of the extra
expense of GM seeds.

Mechanism:

Bt cotton was created through the addition of genes encoding toxin

crystals in the Cry group of endotoxin. When insects attack and eat the
cotton plant the Cry toxins are dissolved due to the high pH level of the
insects stomach. The dissolved and activated Cry molecules bond to
cadherin-like proteins on cells comprising the brush border
molecules. The epithelium of the brush border membranes separates
the body cavity from the gut whilst allowing access for nutrients. The
Cry toxin molecules attach themselves to specific locations on the
cadherin-like proteins present on the epithelial cells of the midge and
ion channels are formed +hich allo+ the flo + of $otassium. @egulat ion
of $otassium concentration is essential and- if left unchec*ed- causes
death of cells. Due to the formation of Cry ion channels su>cient
regulation of $otassium ions is lost and results in the death of e$ithelial
cells. The death of such cells creates ga$s in the #rush #order
mem#rane.

Advantages:
Bt cotton has several advantages over non Bt cotton. The im$ortant
advantages of Bt cotton are #riefly :

• Increases yield of cotton due to effective control of three ty$es of

#oll+orms- viz. American- S$otted and Pin* #oll+orms.

• Insects #elonged to Le$ido$tera (Boll+orms) are sensitive

to crystalline endotoxic $rotein $roduced #y Bt
gene +hich in turn
$rotects cotton from #oll+orms.
• @educt ion in $esticide use in the cultivation of Bt cotton in +hich
#oll+orms are major $ests.

• @educt ion in the cost of cultivation and lo+er farming ris*s.

• @educt ion in environmental $ollution #y the use of insecticides

rarely.

• Bt cotton exhi#it genetic resistance or in#uilt resistance +hich is a

$ermanent ty$e of resistance and not affected #y environmental
factors. Thus $rotects cro$ from #oll+orms.

• Bt cotton is ecofriendly and does not have adverse effect

on
$arasites- $redators- #eneficial insecticides and organisms $resent in
soil.
• It $romotes
multi$lication of
$arasites and $redators
+hich hel$ in
controlling the
#oll+orms #
on larvae and eggs ofy
#oll+orm.
fee
• Go health din
hazards due to rare g
insecticides.
use of

• Bt cotton are early in maturing as com$ared to non Bt cotton.

Disadvantages:
Bt cotton has some limitations

• High cost of Bt cotton seeds as com$ared to non Bt cotton seeds.

• Effectiveness u$ to 756 days- after that the toxin

$roducing e>ciency of the Bt gene drastically reduces.

• Ineffective against suc*ing $ests li*e jassids- a$hids- +hitefly etc.

Bt cotton in India:

Bt cotton is su$$lied in India?s Maharashtra state #y the agri-

#iotechnology com$any- Mahyco- as the distri#utor.

The use of Bt cotton in India has gro+n ex$onentially since its

introduction. @ecent ly India has #ecome the num#er one glo#al ex$orter
of cotton and the second largest cotton $roducer in the +orld. India has
#red Bt-cotton varieties such as Bikaneri Nerma and hy#rids such as
GHH-HH- setting u$ India to #enefit n o + and +ell into the future.

India1s success has #een su#ject to scrutiny. Monsant o?s seeds

are ex$ensive and lose vigour after one generation- $rom$ting the
Indian
Council of Agricultural @esearch to develo$ a chea$er Bt cotton variety
+ i t h seeds that could #e reused. The cotton incor$orated the cry7Ac
gene from the soil #acterium Bacillus thuringiensis (Bt)- ma*ing the
cotton toxic to #oll+orms. In $arts of India cases of ac3uired resistance
against Bt cotton have occurred.

The state of Maharashtra #anned the sale and distri#ution of Bt cotton

in 5675- to $romote local Indian seeds- +hich demand less +ater-
fertilizers and $esticide in$ut- #ut lifted the #an in 567L.

India a$$roved Bt cotton in 5665; n o + it accounts for F5% of all Indian

cotton. Average nation+ide cotton yields + e n t from L65 * gQha in the
5665QL season to a $rojected HD7 * gQha in 5677Q75 J u$ 8F.L%
overall. This chart sho+s the trends in yields- +hich too* off after
Bt +as introduced in 5665. The gra$hs also sho+ that J and here
comes ugly f a c t J in the last H years- as Bt has risen from 67% to
F5% of India1s cotton- yields have dro$$ed steadily.

Biotechnology in
Medicine
Genetically Engineered Insulin ) Hu 0 u l i n *
I n s u l i n is a $e$tide hormone $roduced
#y #eta cells in the $ancreas of various
organisms including human #eings. It
regulates
the 0 e t a b o l i s 0 o / c a r b o h y d r a t e s an
d fats #y $romoting the a#sor$tion
of glucose from the #lood to s*eletal
muscles and fat tissue and #y causing
fat to #e stored rather than used for energy. Insulin also inhi#its the
$roduction of glucose #y the liver.

Exce$t in the $resence of the meta#olic disorder dia#etes

mellitus and meta#olic syndrome- insulin is $rovided + i t h i n the #ody in
a constant $ro$ortion to remove excess glucose from the #lood- +hich
other+ise +ould #e toxic. = h e n #lood glucose levels fall #elo+ a certain
level- the #ody #egins to use stored glucose as an energy source
through glycogenolysis- + hic h #rea*s d o + n the glycogen stored in the
liver and muscles into glucose- +hich can then #e utilized as an energy
source. As a central meta#olic control mechanism- its status is also used
as a control signal to other #ody systems (such as amino acid u$ta*e #y
#ody cells). In addition- it has several other ana#olic effects throughout
the #ody. + h e n c o n t r o l o / i n s u l i n l e 1e l s / ai l s4 d i a b e t e s
0 e l l i t u s can result.

Structure:

Insulin is of t + o
com$osed different $e$tide
chains. ty$esChainof A has 57
aminoand C h a in B has L6 amino
acids
acids. Both chains contain al$ha
helices #ut no #eta strands.
There are L conserved d i s u l f i d e
b r i d g e s +hich hel$ *ee$ the
t + o chains together. Insulin can
also form dimers in solution due
to the hydrogen #onding #et+een the B chains. The dimers can further
interact to form hexamers due to interaction #et+een hydro$ho#ic
surfaces. This scene highlights the hydrophobic and polar parts of an
insulin monomer at a pH of 7.

A number of insulin variants have been made t o favor either the

monomeric or hexameric form. Deletion of the five C terminal residues
of the B chain creates a monomer only form. This portion of the B chain
is involved in hydrogen bonds between the B chain of one monomer and
the A and B chain of another monomer .

Need of Genetically Engineered Insulin:

The original form of the wonder cure for diabetes, these were once the
only type of insulin available, but are now rarely used. A n i m a l i n s u l i n
was originally made
from ground-up
animal pancreas
tissue, and then later
was extracted from
healthy animals
(slaughtered pigs &
cows). The
metabolism of cows and pigs was close enough to human metabolism
that their animal insulin also worked well in human bodies. Beef insulin
has 3 differences from human; pork insulin has 1 difference from
human. The use of a mixture of beef and pork insulin was also possible.
It has been shown that human insulin is less immunogenic than animal
insulin. Porcine insulin is most similar to human insulin. The primary
amino acid sequences of bovine and porcine insulin differ from that of
human insulin by three and one amino acid, respectively. This greater
dissimilarity between human and bovine insulin has been postulated to
be the explanation for the greater antigenicity of bovine insulin as
compared with porcine insulin

One of the p r o b l e m s w i t h a n i m a l i n s u l i n w a s a n t i b o d y i s s ue s . The

body identifies th em and tries to reject them. Pork insulin differs by 1
amino acid and beef insulin by 3 amino acids, so the body's immune
system can sometimes recognize them as foreign.
Immunological complications of insulin therapy have been evident since
animal insulin became available for the treatment of diabetes
mellitus in 1922. In insulin-allergic patients treated with conventional
insulin preparations,
the insulin-s$ecific IgE values are often 76- to 56-fold higher than in
$atients + i t h o u t allergy. It has #een sho+n that human insulin is less
immunogenic than animal insulin. Porcine
insulin is most similar to human insulin. Cross-
reactivity #et + een human insulin and insulin
of animal origin has #een re$orted. A major
$ro#lem is the cross-reactivity that occurs
#et+een anti-insulin anti#odies and the
various animal and human insulin $re$arations
in $atients $resenting + i t h allergy to animal
insulin.

The usage of a n i m a I i n s u I i n h a s s o g r e a t I y d e c I i n e d i n m o d e r n
t i m e s that they have largely #een + i t h d r a + n from the mar * et . Ge+ly
diagnosed dia#etics are ty$ically given synthesized or G e net ica II y
E n g i n e e r e d h u m a n ins uIin .

What is Proinsulin"?

P r o i n s u I i n is the $rohormone $recursor to insulin made in the

#eta cells of t he islets of Langerhans- s$ecialized regions of the
Proinsulin is synthesized on
$ancreas.
mem#rane associated
ri#osomes found on the
endo$lasmic reticulum- +here roughit
is folded and its disulfide
#onds are oxidized. It is
trans$orted then
to the Golgi
a$$aratus + h e r e it is $ac*aged
into secretory vesicles- and
+here it is $rocessed #y a series
of $roteases to form
mature insulin. Mature insulin
has L8 fe+er amino acids; H are
removed
altogether- and the remaining L7 form the C-$e$tide. The C-$e$tide is
a#stracted from the center of the $roinsulin se3uence; the t + o other
ends (the B chain and A chain) remain connected #y disulfide #onds.

= h e n insulin + a s originally $urified from #ovine or $orcine $ancreata-

all the $roinsulin + a s not fully removed. RLRH = h e n some $eo$le used
these insulins- the $roinsulin may have caused the #ody to react + i t h a
rash- to resist the insulin- or even to m a* e dents or lum$s in the s*in at
the $lace +here the insulin + a s injected. This can #e descri#ed as
an iatrogenic injury due to slight differences #et + een the $roinsulin of
different s$ecies. Since the late 7F76s- + he n highly
$urified $orcine insulin + a s introduced- and the level of insulin $urity
reached FF%- this ceased to #e a significant clinical issue. T h e 0 a i n
challenge /or production o/ insulin using rDNA techni6ues 5 a s
getting insulin as s e0bled into 0 a t u r e / o r 0 .

Humulin:

H u 0 u l i n +as the first medication $roduced using modern genetic

engineering techni3ues in + hic h actual human DGA is inserted into a
host cell ( E. coli in this case). Biosynthetic <human< insulin is no+
manufactured for +ides$read clinical use using genetic engineering
techni3ues using recom#inant DGA technology- +hich the
manufacturers claim reduces the $resence of many im$urities- although
there is no clinical evidence to su#stantiate this claim . Eli
L i l l y 0 a r 7 e t e d t h e f i r s t a r t i f i c i a l i nsu l i n4 Hu 0 u l i n 4 i n 8 98 2 .

Humulin $roduction method is as follo+s:

7. DGA coding for A and B $oly$e$tide chains of insulin are

chemically synthesised a in the la#. Sixty three nucleotides are
se3uenced to $roduce A chain of insulin and ninety nucleotide long
DGA designed to $roduce B chain of insulin- $lus terminator codon
is added at the end of each chain se3uence. Anti-codon for
methionine is added a t the #eginning of the se3uence to
distinguish humulin from the other #acterial $roteins.

5. Chemica ly synthesized A and B chain DGA se3uence are inserted

into one of the m a r * e r gene + h i c h are $resent in the $lasmid
vector. Genes are inserted into the $lasmid + i t h the hel$ of
enzymes * n o + n as endonuclease and ligase.

L. The vector $lasmids + it h the insulin gene are then introduced into
the E. coli #acterial cell. These cells are then allo+ed to re$licate
#y mitosis- along + i t h the #acterial cell recom#inant $lasmid also
gets re$licated $roducing the human insulin.

H. A and B $oly$e$tide chains of insulin are then extracted

and
$urified from the fomenters in the la#. High-Performance Li3uid
 Chromatogra$hy (HPLC) is used to get 766% $ure humulin from
the mixture of $roteins.

8. The A and B $oly$e$tide chains of insulin are mixed together and

connected + i t h each other # y disul$hide #ond- forming the
Humulin or synthetic human insulin.

Advantages % Disadvantages of Humulin:

Humulin is the one and only human $rotein $roduced in the #acteria
+ i t h identical chemical structure to that of the natural human insulin.
Administration of humulin reduces the $ossi#ility of anti#ody $roduction
and inflammatory res$onse
in dia#etic $atients. Major
d i > c u l t y is the extraction of
humulin from a mixture of
host $roteins $resent in the
fermentation #roth.

Go+ days to overcome this

extraction $ro#lem synthetic
human insulin are $roduced
in the yeast cell instead of E. coli using the same $rocedure. As yeast is
Eu*aryotes they secrete the +hole humulin molecule + i t h $erfect three
dimensional structures- reducing the need for com$lex and time
consuming $urification methods.

G o+ most of the dia#etic $atients are treated + i t h synthetic human

insulin. Small grou$ of $atients claim that e$isodes of hy$erglycaemic
com$lications have #een increased after shifting from animal origin
insulin to humulin. Go study till date sho+s the difference #et+een the
fre3uency of hy$erglycaemic com$lications in $atient using humulin
(synthetic human insulin) and animal origin insulin.
Gene Therapy
G en e t h e r a p y is the thera$eutic delivery of nucleic acid $olymers into
a $at ient ?s cells as a drug to treat disease. Gene thera$y is an
ex$erimental techni3ue that uses genes to treat or $revent disease. In
the future- this techni3ue
may allo+ doctors to treat a
disorder #y inserting a gene
into a $at ient 1s cells instead
of using drugs or surgery.
@esearchers are testing
several a$$roaches to
thera$y- including: gene

•
@e$lacing a mutated
gene that causes
disease + it h a
co$y of the gene. health
y
•
Inactivating- or / * n o c * i n g out-0 a mutated gene that is functioning
im$ro$erly.

•
Introducing a n e + gene into the #ody to hel$ fight a disease.

Although gene thera$y is a $romising treatment o$tion for a num#er of

diseases (including inherited disorders- some ty$es of cancer- and
certain viral infections)- the techni3ue remains ris*y and is still under
study to m a * e sure that it + ill #e safe and effective. Gene thera$y is
currently only #eing tested for the treatment of diseases that have no
other cures. It should #e noted th at not all medical $rocedures that
introduce alterations to a $at ient ?s genetic ma*eu$ can #e considered
gene thera$y. Bone ma r o + trans$lantation- and organ trans$lants in
general have #een found to introduce foreign DGA into $atients. Gene
thera$y is defined #y the $recision of the $rocedure and the intention of
direct thera$eutic effects.

Gene thera$y +as conce$tualized in 7F75- #y authors + h o urged

caution #efore commencing human gene thera$y studies.

The first attem$t- al#eit an unsuccessful one- at gene thera$y (as +ell
as the first case of medical transfer of foreign genes into humans not
counting organ trans$lantation) + a s $erformed #y Martin Cline on 76
July 7FD6. Cline claimed that one of the genes in his $atients +as active
six months later- though he never $u#lished this data or had it
verified and even if he is correct, it's unlikely it produced any significant
beneficial effects treating beta-thalassemia.

The first germ line gene therapy consisted of producing a genetically

engineered embryo in October 1996. The baby was born on July 21,
1997 and was produced by taking a donor's egg with healthy
mitochondria, removing its nuclear DNA and filling it with the nuclear
DNA of the biological mother - a procedure known as cytoplasmic
transfer.

This procedure was referred to sensationally and somewhat inaccurately

in the media as a "three parent baby", though mtDNA is not the primary
human genome and has little effect on an organism's individual
characteristics beyond powering their cells.

Gene therapy is a way to fix a genetic problem at its source. The

polymers are either expressed as proteins, interfere with protein
expression, or possibly correct genetic mutations.

The most common form uses DNA that encodes a functional,

therapeutic gene to replace a mutated gene. The polymer molecule is
packaged within a "vector", which carries the molecule inside cells.

The first commercial gene therapy, Gendicine, was approved in China in

2003 for the treatment of certain cancers. In 2011 Neovasculgen was
registered in Russia as the first-in-class gene-therapy drug for treatment
of peripheral artery disease, including critical limb ischemia. In
2012 Glybera, a treatment for a rare inherited disorder, became the first
treatment to be approved for clinical use in either Europe or the United
States after its endorsement by the European Commission.

ADA deficiency is one form of SCID (severe combined

immunodeficiency), a disorder that affects the immune system. ADA
deficiency is very rare, but very dangerous, because a malfunctioning
immune system leaves the body open to infection from bacteria and
viruses.
The disease is caused #y a
mutation in a gene on
chromosome 56.
d e f i c i e n c y i s i nADA
herited in
an autoso0al r ec ess i1e
0 a n n e r . The gene codes for
the enzyme adenosine
deaminase (ADA). =ithout
this enzyme- the #ody is
una#le to #rea* d o + n a toxic
su#stance called
deoxyadenosine. The toxin
#uilds u$ and destroys
infection-fighting immune
cells called T and B
lym$hocytes. Because ADA
deficiency affects the
immune system- $eo$le + h o have the disorder are more susce$ti#le to
all *inds of infections- $articularly those of the s*in- res$iratory system-
and gastrointestinal tract. They may also #e shorter than normal. Sadly-
most #a#ies + h o are #orn + i t h the disorder die +ithin a f e + months.

T r e a t 0 e n t s o / ADA D e f i c i e n c y includes:

• #one marro+ trans$lant

gene therapy

• ADA enzyme in PEG vehicle

O n S e p t e 0 b e r 8<4 899=4 t h e f i r s t g e n e t h e r a p y to com#at this

disease +as $erformed #y Dr. = illiam French Anderson on a / ou r >year >
o l d g i r l 4 A s h a n t i DeSil1a- at the Gational Institutes of Health-
Bethesda- Maryland- U.S.A.

Conclusion
Biotechnology is the n e + +onder of science. It is truly multidisci$linary
in nature and it encom$asses several disci$lines of #asic sciences and
engineering. The Science disci$lines from +hich #iotechnology dra+s
heavily are micro#iology- chemistry- #iochemistry- genetics- molecular
#iology- immunology- cell and tissue culture and $hysiology. On the
engineering side it leans heavily on $rocess chemical and #iochemical
engineering since large scale cultivation of microorganisms and cells-
their do+nstream $rocessing are #ased on them. I t c o 0 e s t o u s a s a
g r e a t blessing...
Biotechnology utilizes the techni3ue called genetic engineering or
recom#inant DGA technology +here a microorganism is isolated; its
genetic material is cut- mani$ulated- sealed- again inserted in an
organism and a lo+ed to g r o + in a suita#le environment under
controlled conditions to get the desired $roduct. It loo*s easy #ut is a
very tedious jo# and it ta*es years for a research to achieve its goal.
Li*e every other thing- b i o t e c h n o l o g y t o o h a s s o m e h a r m f u l
impacts:
7. Genetic engineering is a very vital $art of #iotechnology and the
cost of transferring genes fr om one s$ecies to another is very
ex$ensive- + hic h re3uires a huge amount of ca$ital investment.
T he c o s t o f p r o d u c i n g g e n e t i c a l l y - m o d i f i e d p l a n t s a n d
a n i m a l s a r e s k y - r o c k e t i n g and the duration of return are also
not $redicta#le.
5. Genetic engineering crosses #oundaries of re$roduction #y
crossing genes of s$ecies that are com$letely unrelated; hence
giving rise to hazardous results as +ell as also increasing the ris*
of harming multi$le s$ecies.
L. = h e n genetic material from certain viruses is used in the
$roduction of transgenic cro$s- there are chances that these virus
genes + ill com#ine + i t h cro$ genes to $roduce more destructive
viruses. The consum$tion of such cro$s is hazardous to human
health and can cause several life- threatening ailments. It can also
result in cancer- often malignant as +ell.
H. Biotechnology also $oses a num#er of environmental threats.
Genetically modifies cro$s often infect monarch #utteries and
other insect s$ecies.

The a$$lications of #iotechnology are so #road- and the advantages so

com$elling- that virtually every industry is using this technology.
Develo$ments are under +ay in areas as diverse as $harmaceuticals-
diagnostics- textiles- a3uaculture- forestry- chemicals- household
$roducts- environmental cleanu$- food $rocessing and forensics to name
a fe+. Biotechnology is ena#ling these industries to m a* e n e + or #etter
$roducts- often + i t h greater s$eed- e>ciency and flexi#ility.
Biotechnology m u s t continue t o be carefully regulated so t h at
t h e m a x i m u m benefits are received w i t h t h e least risk.

Bi#liogra$hy
http://en.wikipedia.org/biotechnology
http://en.wikipedia.org/insulin
 h t t p : / / w w w. g e n e w a t c h . o r g / s u b -5 6 8 2 3 8
http://en.wikipedia.org/humulin
h t t p : / / w w w. b i o t e c h a r t i c l e s . c o m / O t h e r s -A r t i c l e / H u m a n -

I n s u l i n - a n d - R e c o m b i n a n t - DNA-Te c h n o l o g y - 7 0 . h t m l
https://isaaa.org/resources/publications/pocketk/34/default.

as p
h t t p : / / w w w. s c i e n c e d i r e c t . c o m /
https://en.wikipedia.org/wiki/Gene_therapy
https://en.wikipedia.org/wiki/Adenosine_deaminase_deficie

ncy
h t t p : / / w w w. d i a b e t e s . c o . u k / i n s u l i n / a n i m a l -i n s u l i n . h t m l
B i o l o g y t e x t b o o k (N.C.E.R.T) Class 1 2 t h

Contents

•
Introduction
•
History
•
Biotechnology in Agriculture
Genetically Modified Cro$s
•

@GA Interference (@GAi)

•
 •
Bt toxin
•
Bt cotton
•
Biotechnology in Medicine
•
Genetically engineered insulin
(Humulin)
•
Gene thera$y
•
Conclusion
•
Bi#liogra$hy