BIOLOGY 1

LECTURE 10: CLONING generate force into the bacterial

membrane
CLONING
● As the bacterium divides,
recombinant plasmid is also
DEFINITION Process of producing virtually
replicated & transmitted to the
identical genetic copies of
biological entities progeny bacteria

METHODS Artificial
Natural
Plasmid
● Extrachromosomal DNA found in
IN Process of creating genetically
MOLECULAR identical copies of genes, DNA bacteria
BIOLOGY fragments, cells, or organisms ● Can be used as a cloning vector

Natural Cloning
● Asexual reproduction in prokaryotes
and some plants
● Identical twins in mammals

Identical twins
● Produced when a fertilized egg
splits into two or more embryos
● Contain nearly identical DNA
Clone
● Organism replicated through cloning
● Contains the same genetic makeup
as the original copy
TYPES OF CLONING IN MOLECULAR
BIOLOGY
1. Molecular Cloning
2. Reproductive Cloning
3. Therapeutic Cloning
Molecular Cloning / DNA Cloning
● DNA fragments are joined to
plasmids
● Recombinant plasmid is taken out
of a bacterium via heat/electricity to
Reproductive Cloning / Organismal Cloning
● Creating a new multicellular
organism
● Genetically identical to the source
of the differentiated cell
● Done in mammals via nuclear
transplantation
Nuclear transplantation
● Somatic cell nuclear transfer (SCNT)
● Differentiated cells are obtained
from a donor and cultured in a lab
○ Often mammary cells
● Unfertilized eggs are then obtained
from another donor
● Nucleus from the unfertilized egg is
removed and replaced with the
nucleus of the differentiated cell
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● Newly-replaced egg is grown in a
culture medium and induced to
undergo cleavage
● Early embryo is implanted in a
surrogate mother and allowed to
develop
Dolly the Sheep
● First cloned animal
● Exhibited possible issues with
Reproductive Cloning due to the
emergence of genetic defects
● CC the Cat, ZZ and HH the Monkeys
Cloned animals (transgenic)
● Do not always act, behave, or look
the same as each other
● Many experience genetic defects
○ Acetylation of histones
Epigenetic changes
● Methylation of DNA must be
reversed in the nucleus of the donor
animal so that genes may be
expressed/repressed appropriately
in the early stages of development
● Many genes are turned off in a
normal differentiated cell
○ The affected genes have to
be reprogrammed during the
early stages of development
Therapeutic Cloning
● Creating a cloned embryo for the
production of embryonic stem cells
with the same DNA as the donor cell
○ Also utilizes SCNT
● Can be used in experiments for
creating tissues to replace other
tissues
○ Understanding diseases and
exploring new forms of
treatment
● Risk of rejection by the host
organism is reduced
○ Subject’s own genetic
material is used
○ Cells from another individual
are more likely to be
recognized as foreign
○ Immune system is prompted
to attack “foreign” cells
● Often utilized in agriculture through
plant tissue culture and
micropropagation
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Therapeutic Cloning in Plants
● Plant tissues are removed from the
intact plant in sterile condition
● Tissues are cultured in a neutral
medium
● Single cells suspended in the
medium divide
● Plantlet is cultured on agar medium
then planted in soil

➔ Therapeutic Cloning for plants is a

costly technique
➔ Effective in propagating sterile
plants, plants with small population
sizes, or when seeds cannot be
stored
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LECTURE 11: ASSISTED
REPRODUCTIVE TECHNOLOGY
Assisted Reproductive Technology
● Includes the in vitro handling of
human gametes or embryos for the
purpose of establishing pregnancy
● Used to treat fertility problems for
the purpose of gestational
surrogacy
● Involves surgically removing eggs,
combining them with sperm in a lab,
and returning them to the body or
donating them to another woman
In Vitro Fertilization (IVF)
● Ovaries are stimulated with
hormone therapy to produce eggs
● Eggs and sperm are combined
○ Insemination
○ Intracytoplasmic sperm
injection
● After fertilization, the embryo is
cultured and transferred into the
uterus
○ One or more embryos can be
transferred
● After hormonal treatment, eggs are
retrieved via transvaginal
ultrasound aspiration
Insemination
● Motile sperm are placed together
with oocytes then incubated
overnight
Intracytoplasmic sperm injection (ICSI)
● A single sperm is injected through a
small glass tube into the cytoplasm
of a mature egg cell
● Done in instances of low sperm
count or weak sperm
Laparoscopic oocyte retrieval
● Transabdominal ultrasound-guided
procedure
● Performed when ovaries are
inaccessible through the
transvaginal route
Assisted hatching
● Micromanipulation procedure
● A hole is made in the zona pellucida
prior to embryo transfer
● Helps the fertilized egg hatch out
and attach to uterine wall
Preimplantation genetic diagnosis (PGD)
● Screening for inherited diseases
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● 1-2 cells are removed from the
embryo then tested for specific
genetic diseases
● May be performed prior to embryo
transfer
Ultrasound-guided embryo transfer
● The fertilized egg is transplanted
into the womb with the assistance
of ultrasound imaging
Cryopreservation
● Freezing of remaining embryos for
future transfer
● Can be frozen at the pronuclear
stage (single cell)
○ Or any stage after that
○ Until blastocyst stage (5-7
days after fertilization)
● Also done for individual sperm and
egg cells
Slow freezing
● Embryos are cooled at 3º C per
minute until temperature passes
below -30º C
● Necessitate a Cryopreservation area
○ Liquid nitrogen supply tank
○ Backup embryo freezer
○ Main embryo freezer
○ Storage tanks for frozen
embryos and sperm
Cryoprotectants
● Used to lower the freezing point of
the cryopreservation area
● Help prevent intracellular ice
formation
● Draw water out of the cell via
osmosis
● Cross into the plasma membrane to
replace lost water
COMMON CRYOPROTECTANT
SUBSTANCES
● Ethylene glycol
● Glycerol
● Propylene glycol
Rapid freezing / Vitrification
● Gametes or embryos are exposed to
high concentrations of
cryoprotectants to allow for the
rapid dehydration of cells
● Eggs are loaded into straws which
facilitate ultra rapid cooling
● Straws containing eggs are plunged
into liquid nitrogen at -196º C
Embryo thawing
● Frozen embryos are thawed prior to
transfer into the womb
● When thawed, water is allowed back
into cells and the cryoprotectant is
drawn out
● Performed slowly and in small steps
to prevent cells from fracturing and
dying in the process
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LECTURE 12: REPRODUCTIVE ● A state of complete physical,
HEALTH & SEXUALLY mental, and social well-being in all
matters relating to the reproductive
TRANSMITTED DISEASES
system & its functions and
processes
Sexual health (WHO)
○ Satisfying & safe sex life
● A state of physical, mental, and
○ Capability to reproduce
social well-being in relation to
○ Freedom to decide if, when,
sexuality
and how often to reproduce
● Requires a positive and respectful
○ Right of all to be informed &
approach to sexuality & sexual
have access to safe,
relationships
effective, affordable, and
● Ensures all of the possibility of
acceptable methods of
having pleasurable & safe sexual
family planning
experiences free of coercion,
discrimination, and violence
➔ Sexual health and reproductive
● Necessitates the respect,
health undoubtedly overlap
protection, and fulfillment of the
➔ Both aim to support physiological
sexual rights of all persons
functions
➔ Both seek to reduce the adverse
Sexual health (American SH Association)
outcomes of sexual activity and
● Ability to embrace and enjoy one’s
reproduction
sexuality throughout their own life
● Involves more than sexual behavior
Contraception
● Recognizing and respecting the
● Deliberate prevention of pregnancy
rights shared by all
● Having access to information,
Vasectomy
education, and care
● Surgically cutting off and tying the
● Making an effort to prevent
vas deferens
unintended pregnancies, STDs, and
● Prevents sperm from entering the
to seek care & treatment when
urethra
needed
● Permanent method of contraception
● Being able to experience pleasure,
● Not easily reversible
satisfaction, and intimacy when
desired
Hormonal contraception
● Being able to communicate about
● Birth control pills, injection, patch,
sexual health with others included
vaginal ring
partners & healthcare providers

Reproductive health (WHO)

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● Synthetic estrogen and progestin
are released to block ovulation and
the development of follicles
Abstinence
● Most effective method for
preventing pregnancy
● Not applicable to sexually active
couples
Temporary abstinence
● Natural family planning
● Depends on the framing of sex when
conception is most likely
● Egg can survive in the oviduct for
24-48 hours
● Sperm can survive for 5 days in the
uterus
● A couple practicing this should not
engage in sex for several days
before and after ovulation
Male condom
● Globally most widely used
contraceptive
Female condom
● Latex pouch
● Flexible rubber ring on each end
● One holds in the vagina, one stays
on the outside
● More expensive, but allows women
to take more active and
independent responsibility for
preventing pregnancy
○ In case partner is not willing
Coitus interruptus
● Withdrawal
● Necessitates will power and proper
timing
● Often unsuccessful
Tubal ligation
● Surgically sealing off a section of
each fallopian tube to prevent egg
from traveling to the uterus
Spermicides
● Substances that kill or immobilize
sperm before entering the uterus
● Applied or inserted vaginally prior to
intercourse to prevent pregnancy
● Can be used on its own or combined
with another contraceptive method
Diaphragm
● Silicone cap with flexible ring
● Worn inside the vagina, covering the
cervix
Cervical cap
● Similar to diaphragm but smaller
○ Both tools block sperm from
getting to the egg
● Worn over the cervix
Intrauterine device (IUD)
● Often made of plastic or copper
● Inserted into the uterus to prevent
pregnancy
● Releases copper
○ Via a copper coil
○ Alters the chemistry of the
cervical mucus
○ Makes it more difficult for
sperm to survive
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○ Prevents fertilized eggs from
implanting in the uterine wall
● Protects against pregnancy for 5-10
years
Emergency contraceptive pills (ECP)
● May be used after intercourse
● Disrupt or delay ovulation and
fertilization
○ Can also prevent the
implantation of the embryo
into the uterine wall
Morning-after pills (Plan B)
● Can prevent pregnancy if taken
within 5 days
● Abortifacient
➔ There is no single best
contraceptive method
➔ Different methods suit different
needs and preferences
➔ Religious groups tend to support
only natural contraceptive methods
(abstinence, natural family
planning)
STIs
● May be asymptomatic
● Venereal diseases
STDs
● Commonly spread through sexual
intercourse
● Can also be spread from non-sexual
contact
○ Contaminated blood
○ Breastfeeding
○ During childbirth if mother is
already infected
● Caused by bacteria, viruses, or
parasites
Chlamydia
● Caused by chlamydia trachomatis
● Often asymptomatic
● Can be easily treated with
antibiotics
● Women with chlamydia have an
inflamed cervix but no other
symptoms
○ Sometimes, vaginal
discharge
● In men, inflamed urethra, white
discharge with or without burning
sensation during urination
Gonorrhea
● Gonococcal infection
● Caused by neisseria gonorrhoeae
● Can affect the eyes, mouth, and
throat
○ Also vagina, cervix, uterus,
fallopian tube, urethra, and
anus
● Men - burning when peeing, penile
discharge
○ Pus-like discharge could be
foul-smelling
● In women, asymptomatic half the
time but some have vaginal
discharge and pelvic pain
● Antibiotic medicine will stop
infection, but will not repair any
permanent damage done by the
disease
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○ Antibiotic resistance has
become a problem
○ Successful treatment is
becoming more difficult
Syphilis
● Caused by treponema pallidum
● Primary
○ Firm, painless skin
ulceration
● Second
○ Diffused rashes on the palms
and soles
● Latent
○ Little to no symptoms
● Tertiary
○ Gammas (mass of dead,
swollen fiber-like tissues,
often in the liver)
○ Neurological, cardiac
symptoms
● Can be cured with antibiotics
○ Treatment may not undo any
permanent damage done by
the infection
Genital herpes
● Caused by the herpes simplex virus
(HSV-1, HSV-2)
● Most are unaware that they have
been infected
● May never suffer an outbreak
● Blisters like cold sores
● No cure, but antivirals can shorten
outbreaks
Human immunodeficiency virus (HIV)
● Causative agent of AIDS
○ Acquired immunodeficiency
syndrome
● Infects immune cells
○ cd4+t, helper t-cells,
macrophages, dendritic cells
● Within body fluids, HIV is present as
both free virus particles & viruses
within infected immune cells
● Spread by contact or transfer of
blood, semen, and vaginal fluids
● Can also be transmitted from an
infected mother to infant during
pregnancy, childbirth, or
breastfeeding
● Human body cannot rid itself of the
virus
○ No effective cure
● When infected with HIV, it becomes
a lifetime diagnosis
● Originated from a type of
chimpanzee in central africa
○ Simian immunodeficiency
virus
○ Maybe passed to humans
when they were hunted for
meat & came in contact with
infected blood
○ May have jumped from
chimps to humans as far
back as late 1800s
○ Over the decades, HIV
slowly spread throughout
Africa then other parts of the
world
● The development of AIDS is not
immediate
● If left untreated, only then can HIV
advance into AIDS
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○ Marked by the progressive
failure of the immune AIDS
system ● Most severe
● Badly damaged immune system
Zoonosis ● Vulnerable to infection and
● Diseases passed on from animals to infection-related cancers
humans ○ Opportunistic infections
● Diagnosed when cd4 cell count
STAGES OF HIV drops below 200/mm3 of blood
1. Acute Infection ○ Or develop opportunistic
2. Chronic Infection / Clinical Latency infection
3. AIDS ● High viral load, highly infectious
● Without treatment, most survive for
Acute Infection around 3 years
● Primary HIV infection
● Acute retroviral syndrome Azidothymidine (AZT)
● Occurs 2-4 weeks after initial ● First approved treatment for HIV in
infection 1987
● Large amount of HIV in blood ● Antiretroviral therapy
● Highly contagious ○ Consists of taking various
● Flu-like symptoms HIV medications (HIV
○ Some may not feel sick right treatment regimen) daily
away, if at all ● Prevents HIV from making copies of
● If with flu like symptoms and may itself
have been exposed to HIV, should ● Reduces viral load/amount of HIV in
seek medical care & ask for a test to the body
diagnose acute infection ● With less HIV in body, the immune
● Only antigen antibody/nucleic acid system can recover and produce
tests can diagnose more cd4 cells
○ Even with some in it, the
Chronic Infection immune system can mount
● Asymptomatic HIV infection an attack on infections and
● Still active but reproduces at very certain HIV-related
low levels infections
● May have no symptoms or not get
sick Genital warts
● Without taking medication, may last ● Caused by some types (6, 11) of
a decade or longer human papillomavirus (HPV)
● Can still transmit ● Direct skin to skin contact spreading
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● Often asymptomatic in the early
stages
● Can be treated with the application
of creams, cryotherapy, or surgery
● After treatments, can be resolved in
6 months
○ Sometimes on its own

Trichomoniasis
● Caused by trichomonas vaginalis, a
protozoan parasite
● 70% of women and men do not
have symptoms when infected
● Pain, burning, or itching
● Women may have yellow-green
frothy discharge
● Can be cured with a single dose of
prescription antibiotic medication

Candidiasis
● Fungal infection due to the yeast
candida
● Gential itching and burning
○ Sometimes with white,
cottage cheese-like vaginal
discharge
● Antifungal medication for 3-7 days
usually clears this

➔ Saliva & feces may carry covid

Vertical transmission
● Mother to child transmission during
gestation