Download - Gene Cloning Technology
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GENE CLONING TECHNOLOGY
AND APPLICATION
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I/ADVENT OF GENE CLONING
In the last 1960s:the experimental techniqueswere not sophisticated.
In 1971-1973:A new experimental techniqueswere developed:
Recombinant DNA technology or generticengeneering based on the process of genecloning and the word cloning means identicalcopy
This led to rapid and efficient DNA sequencingtechniques that enabled the structure ofindividual genes tobe determined.
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II/GENE CLONING TECHNOLOGY
What is gene cloning?
Gene cloning is the process in which identical
copies of a particular gene are manufactured by
utilizing molecular biology tools.
Amplified genes are useful in many areas of
research and for medical applications such as
gene therapy.
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Basic steps of gene cloning
1. Target DNA from donororganism extracted,cutwith restrictionendonuclease and ligatedinto a cloning vector cut
with compatiblerestriction endonuclease.
2. Recombinant constructtransferred into host cell.
3. Host cells which do not
take up construct areeliminated by selectionprotocol.
4. Host cell library screenedto identify desired clone ifnecessary.
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Basic steps of gene cloning
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Cloning vectors used for gene cloning
Plasmid vector pBR322 Size:4,36 kb
Carries two antibiotic resistance
genes Tetracycline(Tetr ) and
Ampicillin (Ampr ) This origin allows a
fairly high copy number, about 100copies of the plasmid per cell.
Phage vector
M13mp7 have four
recognize positions of the
resistance Enzyme Highly effective in gene
cloning
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Cosmid cloning system
Lamda cos sites insertedinto a small plasmid
Target DNA ligated
between 2 cosmid DNA
moleculesRecombinant DNA
packaged and E.coli
infected as before
Can clone DNAs up to45kb
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OTHER PLASMID CLONING VECTOR
pUC19 vector palsmidYeast artificial chromosomes(YACs)
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APPLICATION OF GENE CLONING
Medicinal field
This recombinant DNA technology revolution
opens the wide range of methods to solve the
problems in medicine field.This technique is usedto detect the genes which are reponsible for
genetic disorder and this disorder can be corrected
by Gene therapy.
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Gen cloning techniqueis going to acquire vital position to produce many
recombinant Pharmaceuticals like human insulin,recombinant DNA
vaccine,recombinant human growth hormone
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Agricultural fieldGene cloning technology is also securing food position in improving crop
yields,production of herbicides resistant plants,drought resistant plants,disease
resistant plants
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Creating gene library
Gene library of an organism is prepared by gene cloning.
It is a collection of clones sufficient in number to be likely to contain
every single gene present in a particular organism.
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Archeogenetics
An emerging fields known as Archeogenetics is also obtaining benefits
from gene cloning technology.
By examine DNA sequence in living being and fossil,archeologist has
begun to understand the evolutionary origin of modern humans.
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