2. CONTENT:
1)INTRODUCTION
2) RECOMBINANT DNA TECTNOLOGY(Concept)
3) TOOLS USED.
3)IVF ( In-vitro fertilization).
5)HISTORY OF IVF.
6) INDICATION OF IVF.
7)ET( EMBRYO TRANFER).
8) MOLECULAR MAPPING.
9) REFERENCE.
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3. Processes of Recombinant DNA Technology. Recombinant
DNA (rDNA)technology refers to the process of joining DNA molecules
from two different sources and inserting them into a host organism, to
generate products for human use.
TOOLS REQUIRED:-
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4. PLASMID :- pBR322 is 4361 base pairs in length[1] and has two antibiotic
resistance genes – the gene bla encoding
the ampicillin resistance (AmpR) protein, and the gene tetA encoding
the tetracycline resistance (TetR) protein.
It contains the origin of replication of pMB1, and the rop gene, which encodes
a restrictor of plasmid copy number.
The plasmid has unique restriction sites for more than forty restriction
enzymes. Eleven of these forty sites lie within the TetR gene.
There are two sites for restriction enzymes HindIII and ClaI within
the promoter of the TetR gene. There are six key restriction sites inside the
AmpR gene.[2]
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7. INVITRO FERTILIZATION
vitro fertilisation (IVF) is a process of fertilisation where an egg is
combined with sperm outside the body, in vitro (test tube). It involves
monitoring and stimulating a woman's ovulatory process, removing
an ovum or ova (egg or eggs) from the woman's ovaries and letting sperm
fertilise them in a liquid in a laboratory.
After the fertilised egg (zygote) undergoes embryo culture for 2–6 days, it
is implanted in the same or another woman's uterus( sarogate mother),
with the intention of establishing a successful pregnancy.
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8. HISTORY OF IVF-
In 1978 Louise Brown was the first child successfully born after her mother
received IVF treatment. Brown was born as a result of natural-cycle IVF,
where no stimulation was made.
In 1959, M.C.Chang, successfully conducted IVF in rabbits.
February 15, 1969 ,the journal Nature published a paper authored by
R.G.Edwards .B.D.Bavister and P.C.Steptoe; “Early stages of of fertilization
in vitro of human oocytes matured in vitro.
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10. STEPS OF IN-VITRO FERTILIZATION :-
1) INITIAL EVALUATION.
2) SUPPRESSION OF NATURAL HORMONAL CYCLE.
3) COLLECTION OF OOCYTES.
4) COLLECTION OFSPERM.
5) IN VITRO FERTILIZATION OF OOCYTES.
6) EMBRYO TRANSFER(ET)
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11. SUPPRESSION OF NATURAL HORMONAL
CYCLE
Suppression drugs prevent spontaneous ovulation.
In an IVF cycle,it is important that natural ovulation shouldnot occur
(if the eggs leave the ovary ,the doctor will not be able to retrieve them.
Drugs used are as followed;-
ORAL CONTRACEPTIVE PILLS-each day starting starting cycle day 1,2 or3
for approx. 2-4 weeks.
LUPRON/LEUPROLLIDE ACETATE- GnRH agonists; they suppresses LH
surge preventing eggs from being released before the are mature for egg
retrieval .
NAFARELIN- GnRH agonist,administrated 3 times a day by nasal
spray.
CETROTIDE( ganirelix acetate injection)- GnRH agonists.
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12. OVARIAN STIMULATION- it is used to produce
follicles,rather than the single egg normally developed
each month.
SPERM COLLECTION-
Sperm is collected about 60-
90 minutes prior to
fertilization.
Liquefied, centrifuged ,
suspended in culture medium
and incubated for 30-60
minutes at 37*c
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15. Gene mapping is a biotechnological methods used to identify the locus of
a gene and the distances between genes.
The essence of all genome mapping is to place a collection of molecular
markers onto their respective positions on the genome. Molecular
markerscome in all forms.
Molecular markers are used in biotechnology to identify a particular
sequence of DNA in a pool of unknown DNA. A genetic marker may be a
short DNA sequence
Example :-a sequence surrounding a single base-pair change (single
nucleotide polymorphism, SNP), or a long one, like minisatellites.
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19. DROSOPHILLA AS QUEEN OF GENETICS
A fruit fly named Drosophilla Melanogaster posses-
1)short generation time.
2)easy genetic manipulation.
3)large number of progeny.
GENETIC RECOMBINATION:-It is the production of offspring with
combinations of traits that differ from those found in either
parents.
During meiosis in eukaryotes, genetic recombination involves the
pairing of homologous chromosomes.followed by crosing over.
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20. Reference :-
^ "Gene mapping - Glossary Entry". Genetics Home Reference. Bethesda, MD: Lister Hill National Center for
Biomedical Communications, an Intramural Research Division of the U.S. National Library of Medicine.
2013-09-03. Retrieved 2013-09-06.
Brown, Terry A. (2007). Genomes 3. New York, NY: Garland Science Publishing. ISB
"Genetic Mapping Fact Sheet". Bethesda, MD: National Human Genome Research Institute, National
Institutes of Health. Retrieved 2013-09-06.
de La Rochebrochard E, Quelen C, Peikrishvili R, Guibert J, Bouyer J (July 2009). "Long-term outcome of
parenthood project during in vitro fertilization and after discontinuation of unsuccessful in vitro
fertilization". Fertility and Sterility. 92 (1)
van Loendersloot LL, van Wely M, Limpens J, Bossuyt PM, Repping S, van der Veen F (2010). "Predictive
factors in in vitro fertilization (IVF): a systematic review and meta-analysis". Human Reproduction
Update. 16 (6): 577–89.
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