GENE CLONING TECHNOLOGY AND APPLICATION

GENE CLONING TECHNOLOGY

I/ADVENT OF GENE CLONING II/GENE CLONING TECHNOLOGY Definition of gene cloning Basic steps of gene cloning Plasmid vectors used for gene cloning III/APPLICATION OF GENE CLONING Medicinal field Agricultural field Creating gene library Archeogenetics

I/ADVENT OF GENE CLONING

In the last 1960s:the experimental techniques were not sophisticated. In 1971-1973:A new experimental techniques were developed: Recombinant DNA technology or genertic engeneering based on the process of gene cloning and the word cloning means identical copy This led to rapid and efficient DNA sequencing techniques that enabled the structure of individual genes tobe determined.

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.

Basic steps of gene cloning

1. Target DNA from donor organism extracted,cut with restriction endonuclease and ligated into a cloning vector cut with compatible restriction endonuclease. Recombinant construct transferred into host cell. Host cells which do not take up construct are eliminated by selection protocol. Host cell library screened to identify desired clone if necessary.

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Basic steps of gene cloning

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 100 copies of the plasmid per cell.

Phage vector M13mp7 have four recognize positions of the resistance Enzyme Highly effective in gene cloning

Cosmid cloning system

Lamda cos sites inserted into a small plasmid Target DNA ligated between 2 cosmid DNA molecules Recombinant DNA packaged and E.coli infected as before Can clone DNAs up to 45kb

OTHER PLASMID CLONING VECTOR

pUC19 vector palsmid
Yeast artificial chromosomes (YACs)

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 used to detect the genes which are reponsible for genetic disorder and this disorder can be corrected by Gene therapy.

Gen cloning techniqueis going to acquire vital position to produce many recombinant Pharmaceuticals like human insulin,recombinant DNA vaccine,recombinant human growth hormone

Agricultural field
Gene cloning technology is also securing food position in improving crop yields,production of herbicides resistant plants,drought resistant plants,disease resistant plants

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.

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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