DIGESTER GPAT DISCUSSION CENTER : MAKES STUDY EASY

BIOTECHNOLOGY

DIGESTER -77
COMMONLY USED TERMINOLOGIES

S. NO. TERMINOLOGY DEFINITION

1. Transgene The exogenous gene introduced in the target cell.
2. Vector Vehicle or an agent to insert transgene in the defective cell.
3. Virion Mature viral particles
4. Provirus Reverse transcribed DNA of the retrovirus
5. In vivo The process performed directly on the living cell or organism.
6. Ex vivo The process performed on the tissue or cells outside the body.
7. Tropism A biological phenomenon for the growth of an organism.

DIGESTER -78
PLANT CELL AND TISSUE CULTURE MEDIA

TYPE OF
S. NO. FEATURES
CULTURE
1. Callus  Callus culture is the culture of differentiated plant cells
culture induced on media usually containing relatively high auxin
concentrations or a combination of auxin and cytokinin
under in vitro conditions.
 Basal salt mixtures used in plant tissue culture: Murashige
and Skoog medium, White's medium, and woody plant
medium.
 It is used for the synthesis of bioactive secondary metabolites
and for the generation of plants with improved resistance
against salt, drought, diseases and pests.
2. Suspension  A cell suspension or suspension culture is a type of
culture cell culture in which single cells or small aggregates of cells
are allowed to function and multiply in an agitated growth
medium, thus forming a suspension. Suspension
cultures are used in addition to so-called adherent cultures.
 Suspension culture are of 2 types: Batch culture and
Continuous culture.
3. Batch  A large-scale closed system culture in which cells are grown in
culture a fixed volume of nutrient culture medium under specific
environmental conditions (e.g., nutrient type, temperature,
pressure, aeration, etc.) up to a certain density in a tank or
airlift fermenter, harvested and processed as a batch,
especially before all nutrients are used up.

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DIGESTER -79
TYPES OF FERMENTATION

S. NO. TYPE FEATURES INDUSTRIAL USE

1. Lactic acid  Lactic acid is formed from pyruvate  Lactobacillus
fermentation produced in glycolysis. bacteria prepare
 NAD+ is generated from NADH. curd from milk via
 Enzyme lactate dehydrogenase this type of
catalyzes this reaction. fermentation.
 During intense exercise when  Used in the
oxygen supply is inadequate, production of
muscles derive energy by producing sauerkraut.
lactic acid, which gets accumulated
in the cells causing fatigue.
2. Ethanol  Pyruvic acid breaks down into  This is used in the
fermentation acetaldehyde and CO2 is released. industrial
/alcohol  In the next step, ethanol is formed production of wine,
fermentation from acetaldehyde. beer, biofuel, etc.
 NAD+ is also formed from NADH,  In bread and other
utilized in glycolysis. baking process.
 Yeast and some bacteria carry out
this type of fermentation.
 Enzyme pyruvic acid
decarboxylase and alcohol
dehydrogenase catalyse these
reactions.
3. Acetic acid  This is a two-step process.  Vinegar is produced
fermentation  The first step is the formation of by this process.
ethyl alcohol from sugar  Used in textile,
anaerobically using yeast. polymer, paints,
 In the second step, ethanol is further food and beverages
oxidized to form acetic acid using industry.
Acetobacter bacteria. Microbial
oxidation of alcohol to acid is an
aerobic process.
4. Butyric acid  This type of fermentation is  Manufacture of
Fermentation characteristic of obligate anaerobic esters of lower
bacteria of genus Clostridium. alcohols for use as
 It is an important source of energy flavoring agents.
for colorectal epithelium.  Its anhydride is
 Sugar is first oxidized to pyruvate used to make
by the process of glycolysis and then cellulose acetate
pyruvate is further oxidized to form butyrate (CAB), a
acetyl-CoA by the oxidoreductase useful plastic.
enzyme system with the production
of H2 and CO2. Acetyl-CoA is further
reduced to form butyric acid.

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DIGESTER -80
MAJOR INDUSTRIAL ENZYMES FROM BACTERIA, MOLDS AND YEASTS

ENZYME MICROORGANISM APPLICATION

BACTERIAL ENZYMES
Amylase ( and ) • Bacillus  Starch coatings (Paper)
 Desizing (Textiles)
 Removal of stains
 Detergents (Dry cleaning)
Glucose isomerase  Bacillus  Fructose syrup
 Streptomyces
Penicillin amidase • Bacillus • Pharmaceutical formulation
Protease • Bacillus  Detergent
 Spot removing
 Desizing
 Wound cleaning
MOLD ENZYMES
-Amylase • Aspergillus • Baking (Bread)
Glucoamylase  Aspergillus  Syrup and glucose manufacture
 Rhizopus  Digestive aid (Pharmaceutical)
Rennet (aspartic • Aspergillus • Baking
proteinases)
Cellulose  Aspergillus  Liquid, coffee concentrates
 Trichoderma  Digestive aid
 Degradation of wood or wood by-
products
YEASTS
Lipases • Saccharomyces • Food, detergent, and
species pharmaceutical sectors
Dehydrogenase • Saccharomyces  Production of various chemical
species precursors
 Fine chemical industries
Invertase. • Saccharomyces  Hydrolysis of sucrose (Industrial
species production)
 Manufacture of artificial honey,
plasticizing agents used in
cosmetics, pharmaceutical and
paper industries

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DIGESTER -81
ENZYMES USED IN VARIOUS FOOD AND BEVERAGE INDUSTRIES

USAGE ENZYMES
Animal and fish bone Alkaline phosphatase
processing
Baking -Amylase, Protease, Phospholipases, Xylanase
Brewing production of beer Amylases (- and -), Protease, Cellulases, Papain,
and alcoholic beverage Aminoglucosidases, Xylanase
Dairy Chymosins (animal/microbial), Lipase, Lactase, Lysozyme
Egg white processing Glucose oxidase, Catalase
Fruit and vegetable Pectin esterase, Pectin lyase, Hemicellulases,
processing Polygalacturanase
Meat processing Protease, Papain
Starch and sugar Amylase (- and -), Glucoamylase, Pollulanase, Invertase,
Glucose Isomerase, Xylanase

DIGESTER -82
TYPES OF RESTRICTION ENDONUCLEASES
TYPE FEATURES
Restriction  A single enzyme with 3 subunits for recognition,
Endonucleases I cleavage and methylation. It can cleave up to 1000 bp
from recognition site.
 Type I enzymes are complex, multi-subunit, combination
restriction-and-modification enzymes that cut DNA at
random far from their recognition sequences.
Restriction  Type II enzymes cut DNA at defined positions close to or
Endonucleases II within their recognition sequences.
 They produce discrete restriction fragments and distinct
gel banding patterns.
 They are the only class used in the laboratory for
routine DNA analysis and gene cloning.
 It includes sub-classes i.e., Type IIF, Type IIE, Type IIM,
Type IIG, Type IIS, etc.
Restriction  A single enzyme with 2 subunits for recognition and
Endonucleases III cleavage.
 Type III restriction enzymes (e.g., EcoP15) recognize two
separate non-palindromic sequences that are inversely
oriented.
 They cut DNA about 20–30 base pairs after the
recognition site.
 They are components of prokaryotic DNA restriction-
modification mechanisms that protect the organism
against invading foreign DNA.

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DIGESTER GPAT DISCUSSION CENTER : MAKES STUDY EASY

Restriction  Type IV enzymes recognize modified, typically

Endonucleases IV methylated DNA and are exemplified by the McrBC and
Mrr systems of E. coli.
Restriction  Type V restriction enzymes (e.g., the cas9-gRNA complex
Endonucleases V from CRISPRs) utilize guide RNAs to target specific non-
palindromic sequences found on invading organisms.
 They can cut DNA of variable length, provided that a
suitable guide RNA is provided.
Artificial restriction  Artificial restriction enzymes can be generated by
enzymes fusing a natural or engineered DNA binding domain to
a nuclease domain.
 Such artificial restriction enzymes can target large DNA
sites (up to 36 bp) and can be engineered to bind to
desired DNA sequences.
 Zinc finger nucleases are the most commonly used
artificial restriction enzymes and are generally used
in genetic engineering applications, but can also be
used for more standard gene cloning applications.

DIGESTER -83
SOURCES OF DNA FOR CLONING

S. NO. SOURCE FEATURES

1. Amplified DNA The polymerase chain reaction (PCR) is a relatively
simple technique that amplifies a DNA template to
produce specific DNA fragments in vitro.
2. cDNA In genetics, complementary DNA (cDNA) is DNA
synthesized from a single-stranded RNA (e.g.,
messenger RNA (mRNA) or microRNA (miRNA))
template in a reaction catalyzed by the enzyme reverse
transcriptase.
cDNA is often used to clone eukaryotic genes in
prokaryotes.
3. Genomic DNA Genomic DNA, or gDNA, is the chromosomal DNA of
an organism, representing the bulk of its genetic
material.
4. Synthetic DNA Synthetic DNA constructs are designed and
manipulated using computer-aided design software.

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DIGESTER -84
GENE THERAPY VECTORS

S. NO. CLASS TYPES FEATURES

1. Viral Retrovirus The retrovirus is an RNA virus, the genome of it is
vectors made up of the RNA (not DNA). It has two RNA
molecules in their genome. Therefore, it is also
known as retrovirus-mediated gene therapy.
 Stable integration
 Infect replication cellsSuitable for ex
vivo treatment
Lentivirus The lentivirus is another form of the retrovirus
that can even infect the non-dividing cells.
HIV is one of the best examples of Lenti-
retrovirus.
 Infect proliferating, non-proliferating and bone
marrow cells.
 Self-inactive
Adenovirus Adenovirus vectors are the most commonly
employed vector for cancer gene therapy. They are
also used for gene therapy and as vaccines to
express foreign antigens.
 Infect both dividing as well as non-dividing
cells.
 Chance of infection is less
Adeno Adeno-associated virus (AAV) is a non-enveloped
associated virus that can be engineered to deliver DNA to target
virus cells.
 Longer transgene expression.
 Non-pathogenic
 Broad tropism
2. Non-viral Liposome The liposome also called lipoplex-mediated gene
vector therapy is an artificial technique non-infected to
the host cell. The liposomes are artificially
synthesized molecule typically 0.025 to 2 μm in
size, made up of the lipids.
Transposon The transposons are mobile genetic
elements that can move from one location to
another into the genome. It also contains coding
genes and terminal repeats as like the viruses.
Naked DNA Naked DNA refers to DNA that is not associated
with proteins, lipids, or any other molecule to help
protect it.

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DIGESTER -85
DNA VECTORS

S. NO. TYPES FEATURES

1. Plasmids • A plasmid is a small, extrachromosomal DNA
molecule within a cell that is physically separated
from chromosomal DNA and can replicate
independently.
• Plasmids are the most-commonly used bacterial
cloning vectors. These cloning vectors contain a site
that allows DNA fragments to be inserted.
2. Viral vectors • Viral vectors are tools commonly used by
molecular biologists to deliver genetic
material into cells. This process can be performed
inside a living organism (in vivo) or in cell culture (in
vitro). Ex. - Bacteriophage λ, M13, etc.
3. Cosmids & • A cosmid is a type of hybrid plasmid that contains
Phagemids a Lambda phage cos sequence.
• They are often used as a cloning vector in genetic
engineering.
• A phagemid or phasmid is a DNA-based cloning
vector, which has both bacteriophage and plasmid
properties. These vectors carry, in addition to the
origin of plasmid replication, an origin of replication
derived from bacteriophage.
4. Artificial • Artificial chromosomes are cloning vectors that can
chromosomes carry DNA inserts orders of magnitude larger than is
possible with plasmids or lambda-phage-derived
vectors.
• Examples: -
 BAC - Bacterial artificial chromosome
 YAC - Yeast artificial chromosomes
 MAC - Mammalian artificial chromosome

DIGESTER -86
METHODS OF GENE TRANSFER

S. NO. METHOD FEATURES

1. Transformation It is the method by which genetic material in the form
of “naked” deoxyribonucleic acid (DNA) is
transferred between microbial cells.
2. Conjugation Conjugation is defined as the transfer of DNA in a
site- and strand-specific manner from a donor to a
recipient cell, which have formed close contacts with
one another, that is, a mating pair.

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3. Electroporation Electroporation is a physical transfection method

that uses an electrical pulse to create temporary pores
in cell membranes through which substances like
nucleic acids/gene/drug can pass into cells.
4. Liposome-mediated Lipofection (or liposome transfection) is a
gene transfer technique used to inject genetic material into a cell by
(Lipofection) means of liposomes, which are vesicles that can easily
merge with the cell membrane since they are both
made of a phospholipid bilayer.
5. Transduction Transduction, a process of genetic recombination in
bacteria in which genes from a host cell (a bacterium)
are incorporated into the genome of a bacterial virus
(bacteriophage) and then carried to another host cell
when the bacteriophage initiates another cycle of
infection.
6. Direct transfer of The DNA are direct transferred into the cell nucleus by
DNA Microinjection and particle bombardment
techniques.

DIGESTER -87
RECOMBINANT PROTEINS APPROVED FOR HUMAN USE

RECOMBINANT
S. NO. PROTEIN COMPANY INDICATION
PRODUCT
1. Insulin Humulin Eli Lilly Diabetes
lnsulin Human AF Novo Nordisk mellitus
2. Erythropoietin EPOGEN/Procrit Amgen, Ortho Biotech Anemia
3. Human Growth Somatropin/ Eli Lilly, Genetech, Dwarfism
Hormone Protropin/ Upjohn, Novo Nordisk
Humatrope
4. Factor VIII ADVATE/Kogenate Baxter, Bayer Hemophilia A
5. Factor IX BeneFix Genetics Institute Hemophilia B
6. DNase I Dornase alfa Genetech Cystic fibrosis
(Pulmozyme)
7. Tissue Activase/Alteplase Genetech Acute
plasminogen myocardial
activator (TPA) infarction
8. -Interferon Intron A Schering-Plough Hairy cell
corporation leukemia
9. Hepatitis-B Recombivax HB / Merck Research Hepatitis B
vaccine Engerix B Laboratories

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DIGESTER -88
HUMAN GENE THERAPY TRIALS

S. NO. DISEASE GENE THERAPY

1. AIDS rev and env
2. Breast cancer Multidrug resistance I
3. Citrullinemia Arginosuccinate synthetase
4. Colorectal cancer, Histocompatibility locus antigen-B7
Melanoma, Renal cancer (HLA-B7)
5. Cystic fibrosis Cystic fibrosis transmembrane
regulator (CFTA)
6. Diabetes Glucose transporter-2 (GLUT-2),
Glucokinase
7. Duchenne muscular Dystrophin
dystrophy
8. Emphysema α1-Antitrypsin
9. Familial Low density lipoprotein (LDL) receptor
hypercholesterolemia
10. Fanconi anemia Fanconi anemia C
11. Gaucher's disease Glucocerebrosidase
12. Glioblastoma (brain Thymidine kinase (herpes simplex
tumor), AIDS, Ovarian virus)
cancer
13. Head and neck cancer P53
14. Hemophilia B Factor IX
15. Lesch-Nyhan syndrome Hypoxanthine-guanine phosphoribosyl
transferase (HGPRT)
16. Melanoma Tumor necrosis factor (TNF)
17. Melanoma, Renal cancer Interleukin-2 (IL-2)
18. Peripheral artery Vascular endothelial growth factor
disease (VEGF)
19. Phenylketonuria Phenylalanine hydroxylase
20. Severe combined Adenosine deaminase (ADA)
immunodeficiency
(SCID)
21. Short stature Growth hormone
22. Sickle-cell anemia β-Globulin
23. Thalassemia α- or β-Globulin

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DIGESTER -89
ENZYMES USED IN RECOMBINANT DNA TECHNOLOGY/GENETIC ENGINEERING

S. NO. ENZYME USE/REACTION

1. Alkaline Removes phosphate groups from 5’-ends of double
phosphatase single/stranded DNA (or RNA)
2. Bal 31 nuclease For the progressive shortening of DNA
3. DNA ligase Joins DNA molecules by forming phosphodiester linkages
between DNA segments
4. DNA polymerase I Synthesizes DNA complimentary to a DNA template
5. DNase I Produces single-stranded nicks in DNA
6. Exonuclease III Removes nucleotides from 3’-end of DNA
7. Polynucleotide Transfers phosphate from ATP to 5’-OH ends of DNA or
kinase RNA
8. Restriction Cut double-stranded DNA with a specific recognition site
enzymes
9. Reverse Synthesizes DNA from RNA
transcriptase
10. RNase A Cleaves and digests RNA (and not DNA)
11. RNase H Cleaves and digests the RNA strand of RNA-DNA
heteroduplex
12. SI nuclease Degrades single-stranded DNA and RNA
13. Taq DNA Used in polymerase chain reaction (PCR)
polymerase
14. Terminal • Adds nucleotides to the 3'-ends of DNA or RNA
transferase • Useful in homopolymer tailing
15. λ exonuclease Removes nucleotides from 5’-end of DNA

DIGESTER -90
MAJOR TYPES OF FILTRATION PROCESSES

SIZE OF PARTICLES COMPOUND OR PARTICLE

TYPE
Microfiltration
Ultrafiltration
Reverse osmosis
(Hyperfiltration)
SEPARATED SEPARATED
0.1 – 10 m Cells or cell fractions, viruses.
0.001-0.1 m Compounds with molecular weights
greater than 1000 (e.g., Enzymes).
0.0001-0.001 m Compounds with molecular weights
less than 1000 (e.g., Lactose).

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DIGESTER -91
TRANSGENIC ANIMALS AS BIOREACTORS FOR THE PRODUCTION OF PROTEINS

TRANSGENIC ANIMAL PROTEIN PRODUCT BIOLOGICAL IMPORTANCE

Cow Lactoferrin  Promotes intestinal iron
absorption and hence can be
used to overcome iron-deficiency
anemias.
 Possesses antibacterial activity.
Cow Interferon  Provides resistance against viral
infections
Goat Cystic fibrosis  For the treatment of patients
transmembrane suffering from cystic fibrosis
regulator (CFTR) (promotes transport of ions).
Goat Tissue plasminogen  Used in treating the patients of
activator (tPA) myocardial infarction (dissolves
blood clots).
Goat Antithrombin III  Regulates blood clotting
Goat and other Vaccines  To immunize against various
animals diseases
Mouse Urokinase  For dissolving blood clots
Mouse Immunoglobulins  Administration enhances
(antibodies) immunity
Pig Hemoglobin  Blood transfusion
Rabbits -Glucosidase  Treatment of Pompe’s disease (a
genetic disorder characterized by
block in glycogen degradation).
Sheep 1- Antitrypsin  Used in the treatment of
emphysema.
 Promotes the exchange of gases
in lungs

DIGESTER -92
rPROTEINS APPROVED FOR HUMAN USE BY USFDA

S. NO. RECOMBINANT PROTEIN DISORDER (S) TREATED

1. Coagulation factor IX Christmas disease/Hemophilia B
2. Coagulation factor VIII Hemophilia A
3 DNase 1 Cystic fibrosis
4. Erythropoietin Anemia,
Kidney disease
5. Glucocerebrosidase Gaucher's disease
6. Granulocyte colony Cancer
stimulating factor
7. Growth hormone Growth defects in children

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8. Hepatitis B surface antigen Hepatitis B

(vaccine)
9. Insulin Diabetes mellitus
10. Interferon α Leukemia, Kaposi sarcoma genital warts,
Hepatitis B
11. Interferon β Multiple sclerosis
12. Interferon γ Chronic granulomatous disease
13. Interleukin-10 (IL-10) Thrombocytopenia
14. Interleukin-2 (IL-2) Renal cell carcinoma
15. Somatotropin Growth defects
16. Tissue plasminogen Acute myocardial infarction, Pulmonary
activator (tPA) embolism

DIGESTER -93
RECOMBINANT PROTEINS FOR TREATMENT OF HUMAN DISORDERS

S. NO. DISORDER RECOMBINANT PROTEIN (S)

1. Anemia Hemoglobin, Erythropoietin
2. Asthma Interleukin-I receptor
3. Atherosclerosis Platelet derived growth factor
4. Delivery Relaxin
5. Blood clots Tissue plasminogen activator, Urokinase
6. Burns Epidermal growth factor
7. Cancer Interferons, Tumor necrosis factor, Colony
stimulating factors, Interleukins,
Lymphotoxin, Macrophage activating
factor
8. Diabetes Insulin, Insulin-like growth factor
9. Emphysema α1-Antitrypsin
10. Female infertility Chorionic gonadotropin
11. Free radical damage (minimizing) Superoxide dismutase
12. Growth defects Growth hormone, Growth hormone-
releasing factor, Somatomedin-C
13. Heart attacks Prourokinase
14. Hemophilia A Factor VIII
15. Hemophilia B Factor IX
16. Hepatitis B Hepatitis B vaccine
17. Hypoalbuminemia Serum albumin
18. Immune disorders Interleukins, β-cell growth factors
19. Kidney disorders Erythropoietin
20. Low Gehrig's disease Brain-derived neurotropic factor
(amyotrophic lateral sclerosis)
21. Multiple sclerosis Interferons (α, β, γ)
22. Nerve damage Nerve growth factor
23. Osteomalacia Calcitonin
24. Pain Endorphins and Enkephalins
25 Rheumatic disease Adrenocorticotropic hormone
26. Ulcers Urogastrone
27. Viral infections Interferons (α, β, γ)

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DIGESTER -94
ANALYTICAL METHODS FOR BIOTECHNOLOGY PROCESS VALIDATION

S. NO. METHODS DETECTION

1. Bioassays • Potency
• Tertiary structure of proteins
2. Carbohydrate analysis • Glycoforms,
• Carbohydrate sequence
3. Electrophoresis • Purity,
• Impurities,
• Glycoforms
4. HPLC • Purity and impurities,
• Carbohydrate analysis
5. Mass spectrometry • Purity and impurities,
• Molecular weight,
• Glycosylation
6. Nucleic acid sequencing • Genetic stability
7. Polymerase chain • DNA,
reaction • Viruses,
• Mycoplasm
8. Peptide mapping • Impurities
9. Western blot • Protein impurities

DIGESTER -95
TYPES OF IMMUNE RESPONSES

PRIMARY IMMUNE RESPONSE SECONDARY IMMUUNE RESPONSE

Slow & short-lived Prompt, powerful and prolonged
Log phase - Long Log phase - Short
Low- Titer of antibody High titer of antibody
IgM is secreted IgG is produced
No negative phase Negative phase may be seen
Produced after one dose Produced after multiple doses
Level of antibody is not effective Level of antibody is effective

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DIGESTER -96
TYPES OF IMMUNITY

I. INNATE (NON-SPECIFIC) IMMUNITY

1. First line of defense
 Intact skin
 Mucous membranes and their secretions
 Normal microbiota
Physical factors
i. Epidermis of skin Forms a physical barrier to the entrance of microbes.
ii. Mucous Inhibit the entrance of many microbe but not as effective as
membranes intact skin.
iii. Mucus Traps microbe in respiratory and gastrointestinal tracts.
iv. Lacrimal Tears dilute and wash away irritating substances and
apparatus microbes.
v. Saliva Washes microbes from surfaces of teeth and mucous
membranes of mouth.
iv. Hairs Filter out microbes and dust in nose
iiv Cilia Together with mucus, trap and remove microbes and dust from
upper respiratory tract.
iiiv Epiglottis Prevents microbes from entering lower respiratory tract.
ix. Urine Washes microbes from urethra.
x. Vaginal secretion Move microbes out of female reproductive tract.
xi. Defecation and Expel microbes from body.
vomiting
Chemical factors
i. Sebum Forms a protective acidic film over the skin surface that
inhibits growth of many microbes.
ii. Lysozyme Antimicrobial substance in perspiration, tears, saliva, nasal
secretions and tissue fluids.
iii. Gastric juice Destroys bacteria and most toxins in stomach.
iv. Vaginal secretions Slight acidity discourages bacterial growth.
2. Second line of defense
 Natural killer (NK) cells and phagocytic white blood cells.
 Inflammation
 Fever
 Antimicrobial substances
Defensive cells
i. Phagocytes Phagocytosis by cells such as neutrophils and macrophages.
ii. Natural killer (NK) Kill infected target cells by releasing granules that contain
cells perforin and granzymes. Phagocytes then kill the infected
microbes
iii. Inflammation Confines and destroy microbes and initiates tissue repair.
iv. Fever Intensifies the effects of interferons, inhibits growth of some
microbe and speeds up body reactions that aid repair.

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Antimicrobial substances
i. Complement Causes cytolysis of microbes, promotes phagocytosis and
system contributes to inflammation.
ii. Interferons Protect uninfected host cells from viral infection.
iii. Transferrins Inhibit growth of certain bacteria by reducing the amount of
available iron.
iv. Antimicrobial Cause lysis of bacteria
peptides
II. ADAPTIVE (ACQUIRED) IMMUNITY
3. Third line of defense
 Specialized lymphocytes: B cells and T cells
 Antibodies
Naturally Acquired Artificial Acquired
Active Passive Active Passive
Antigens enter the Antibodies pass Antigens are Preformed
body naturally; body from mother to introduced in antibodies in
induces antibodies & fetus via placenta or vaccines; body immune serum are
specialized to infant via the produces antibodies introduced by
lymphocytes. mother’s milk. & specialized infection.
Example - Wild Example - The lymphocytes. Example - The
infection i.e., with placental transfer Example – rabies vaccine and
Hepatitis A virus of IgG from mother Vaccination - A snake antivenom.
(HAV) and to fetus during vaccine stimulates
subsequent recovery pregnancy that a primary
gives rise to generally lasts 4 to 6 response against
a natural active months after birth the antigen without
immune response The IgA and IgG causing symptoms
usually leading to found in human of the disease.
lifelong protection. colostrum.

DIGESTER -97
TYPES OF HYPERSENSITIVITY REACTIONS

Type I (Anaphylactic)  Occurs within minutes of exposure to antigen.

reactions  An antigen combines with IgE antibodies.
 IgE Binds to most cells and Basophils causing them to
undergo degranulation and release several mediators.
 Histamine: Dilates and increases permeability of Blood
vessels, increases mucus secretion, smooth muscle
constriction.
 Prostaglandins: Contraction of smooth muscle of
Respiratory system and increases mucus secretion.
 Leukotrienes: Bronchial spasms
 Massive drop in blood pressure.

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 DIGESTER : BIOTECHNOLGY DIGESTER

Type II  Involvement activation of complement by IgG or IgM binding

(Cytotoxic) reactions to an antigenic cell. Antigenic cell is lysed.
 Transfusion reactions - Incompatible donor is lysed as they
enter blood stream.
 Rh system: 85% of population is Rh +ve. Those who are Rh-
ve can sensitized to destroy Rh positive blood cells.
 Hemolytic disease of Newborn: Fetal Cells are destroyed by
maternal anti-Rh antibodies that cross the placenta.
Type III  Involves reaction against soluble antigens circulating in
(Immune complex) serum usually involves IgA antibodies.
reactions  Antibody-Antigen Immune complexes are deposited in
organs, active complement and causes inflammatory kidney
damage.
 Glomerulonephritis: Inflammatory kidney damage.
 Rheumatoid arthritis, systemic lupus erythematosus
Type IV (Cell  Reactions are delayed by one or more days (delayed type
mediated) reactions hypersensitivity).
 Delay is due to migration of macrophages and T-cells to site
of foreign antigens.
 Reactions are frequently dispatched on skin.

DIGESTER -98
CHARACTERISTIC OF HYPERSENSITIVITY REACTIONS

TYPES Type –I Type-II Type-III Type- IV

(Anaphylactic) (Cytotoxic) (Immune complex) (Delayed type)
Characteristic

Antigen Exogenous Cell surface Soluble Tissues and

organs
Antibody IgE IgG, IgM IgG, IgM None
Appearance Wheal and Lysis and Erythema, edema Erythema and
flare necrosis and necrosis induration
Histology Basophils and Antibody and Complement and Monocytes and
Eosinophils complement neutrophils lymphocytes
Response time 15-30 minutes Minutes- 3-8 hours 48-72 hours
hours
Transferred Antibody Antibody Antibody T-cells
with
Examples Allergic asthma, Erythroblast SLE, Farmer’s lung Tuberculin test,
Hay fever osis fetalis, disease Poisoning,
Goodpasture Granuloma
syndrome,
Nephritis

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DIGESTER GPAT DISCUSSION CENTER : MAKES STUDY EASY

DIGESTER -99
PATTERNS OF INHERITENCE AND GENETIC DISORDERS

PATTERN OF
S. NO. FEATURES GENETIC DISORDERS
INHERITANCE
1. Autosomal • Autosomal dominant  Myotonic muscular
Dominant inheritance is a way a genetic trait dystrophy
or condition can be passed down  Huntington disease
from parent to child.  β-Thalassemia
• One copy of a mutated gene from  Familial
hypercholesterolemia
one parent can cause the genetic
 Familial
condition.
retinoblastoma
• A child who has a parent with the
mutated gene has a 50% chance of
inheriting that mutated gene.
2. Autosomal • This genetic condition can occur  Sickle cell anemia
Recessive when the child inherits one copy of  Cystic fibrosis
a mutated gene from each parent.  Phenylketonuria
The parents of child with an  Tay-Sachs disease
autosomal recessive condition  Severe Combined
Immunodeficiency
usually do not have the condition.
Disease (SCID)
• Unaffected parents are called
carriers because they each carry
one copy of the mutated gene and
can pass it to their children.
3. X-chromosome • X-linked inheritance means that  Color blindness
(Sex the gene causing the trait or the  Hemophilia (A/B)
chromosome)- disorder is located on the X  Muscular dystrophy
linked chromosome.
inheritance • Females have two X
chromosomes while males have
one X and one Y chromosome.
4. Mitochondrial • The inheritance of a trait encoded  Leber's hereditary
chromosome in the mitochondrial genome. optic neuropathy
linked • Persons with a mitochondrial (LHON)
inheritance disease may be male or female but  Chronic progressive
they are always related in the external
ophthalmoplegia
maternal line and no male with the
(CPEO)
disease can transmit it to his
children.

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 DIGESTER : BIOTECHNOLGY DIGESTER

DIGESTER -100
TYPES OF ENZYME IMMOBILIZATION

COVALENT MEMBRANE
CHARACTERISTICS ADSORPTION ENTRAPMENT
BONDING CONFINEMENT
Involved bonds of Weak force Covalent bonds Physical Spherical
Process such as Van der mainly, others entrapment in particle
Walls force and sulphide, oxide, gel or fiber formation within
hydrogen bond amino, semi-permeable
carboxyl, membrane
hydroxyl,
ammonium,
phenol ring
Factor affecting pH, Ionic Not affected by pH, pH,
immobilization strength, general Temperature, Temperature,
Temperature conditions addition of addition of
solvents solvents
Carrier matrix Calcium- DEAE- cellulose, Polyacrylamide Semipermeable
Phosphate, polyurethane gel, collagen, membrane via
Charcoal, gelatin, starch, hollow fibers,
Cellulose, cellulose, liposomes
DEAE- silicon and
Sephadex rubber
Enzymes -amylase, -amylase, Amino cyclase Amino cyclase
Catalase, Cellulose,
Invertase Pectinase
Advantages Cost factor is Quite simple Simple and Highly effective
low so can be and cost effective method
used in small effective
level industry
Disadvantages Enzyme leakage Denaturation of Leakage from Cost factor is
during usage enzyme by the the matrix high, problems
polyfunctional encountered
reagents during operation
is high

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