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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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