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   RECOMBINANT DNA TECHNOLOGY

 

 

 

 

INTRODUCTION

 

 

       A recombinant DNA molecule is produced by joining together two or more DNA segments usually originating from different organisms. More specifically a recombinant DNA molecule is a vector into which the desired DNA fragment has been inserted to enable its cloning in an appropriate host. This is achieved by using specific enzymes for cutting the DNA into suitable fragments and then for joining together the appropriate fragments. In this manner a recombinant DNA molecules maybe produced which contain a gene from in organism joined to regulatory sequences from another organisms, such a gene is called chimerical gene. Clearly, the capability to produce recombinant DNA molecule has given man the power and opportunity to create novel gene combinations to suit specific needs. Recombinant DNA molecule are proceed with one of the following three objectives (1) to obtain a large number of copies of specific DNA fragment (2) to integrate the gene in question into the chromosome of a produced by the concerns gene (3) to  integrate the gene in question into the chromosome of a target organism where it expresses itself. Even fort the latter two objectives, it is essential to first obtain large number of copies of the concerned genes. To achieves this, NA segments are integrate in to a self replication DNA molecule called vector, most commonly used vectors are either bacterial plasmids or DNA  viruses. All these steps concerned with piecing tighter DNA segments of diverse origin and placing them into a suitable vector together constitute recombinant DNA technology.

 

      The vectors containing DNA segments to be cloned, called DNA inserts, are then introduced into a suitable organism, usually a bacterium, this organism is called host, while the process is called transformation. The transformed host cells are selected and cloned. The recombinant DNA present in such cloned would replicate either in synchrony with or independent of the host cell the gene present in the vector may or may not express itself by directing the synthesis of concerned polypeptide .The step concerned with transformation of a suitable host with recombinant DNA and cloning of the transformed cell is called DNA cloning or gene cloning. However, often DNA or gene cloning is taken to include both the development of recombinant DNA as well as their cloning in a suitable host.

 

     

 

RECOMBUNANT DNA TECHNOLOGY IN BRIEF

 

 

     Recombinant DNA technology is a major field of biotechnology witch is completed in a deep description. There are many several points to study Recombinant DNA technology in briefly.-

 

     Introduction to gene cloning,. Tools and enzymes used inn gene manipulation, restriction enzyme DNA ligase, DNA polymerase, reverse transcriptase, polynucleotide kinase, end labeling and other process used for DNA technology. Major cloning vehicle and their application plasmid vector, phagemid vectors, YAC, BACX, Ti plasmid, expression vectors, shuttle vectors, binary vectors, transposones. Making of genomic and cDNA libraries, their screening and major application. Production of transgenic microbes and their application in biotechnology. Production of transgenic animals and their application in biotechnology. Requirement of recombinant molecule in health, pharmaceuticals, agriculture, and industrial sectors, in search labs. Rationals for the design of vectors, over expression of the recombinant proteins, ribosome binding sites, purification tags, protease cleavage sites, and inducible expressing system. Over expression system, production of inclusion body. Determination of purity and activity of over expressed proteins.

 

 

CONCLUSION AND FUTURE PROSPECTS

 

 

      Recombinant DNBA technology has made the transfer of genes from any organism to any other organism a technical feasibility, initially, genes of interest from eukaryotes were transferred into prokaryotes like E.coli to obtain useful an valuable biochemical’s. An interesting plant gene used in such transfer is that producing thaumatin isolated from the African shrub thaumatococcus daniellii. Thaumatin is a protein that is 100,000 times sweeter than sugar on molar bases. And has flavor enhancing effect at sub sweetness. The gene producing thaumatin has been transferred and expressed in E. coli yeast, in an attempt to obtain thaumatin at commericially acceptable prices, the cost of production, however, remains unacceptably high due to rather low levels of expression.

 

     The second phase in gene transfers concerned with the mobilization of genes from prokaryotes and eukaryotes into plants to afford protection from various biotic and abiotic stresses. The third and current phase of gene transfers aims at placing genes in plants with a view to produce valuable biochemicals. Transgenic plants. Offer several unique advantages in biochemical production and storage over their production through bacterial fermentation. However, the cost of isolation and purification of these biochemical’s from plants makes them commercially scale from transgenic plants, further refinements in downstream processing produces are expected to make it commercially attractive to grow crops for harvesting novel biochemical’s in addition to/in place of the conventional produce of the concerned crop.

 

 

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