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Transformation, Transduction and Transfection –Gene transfer methods
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Gene transfer is a technique to stably and efficiently introduce functional genes (that are usually cloned) into the target cells. Genes are the fundamental hereditary units of all the life forms. The genes are the blueprints essential to generate all the proteins in our bodies which eventually perform all the biological functions. Therefore, when a gene is efficiently introduced into a target cell of the host, the protein which is encoded by that gene is produced.

Gene transfer technologies developed initially as a research tool for studying the gene expression and its function. On the other hand, as novel gene transfer technologies developed and older technologies were sophisticated, its potential applications have expanded significantly. A range of techniques and naturally occurring processes are utilized for the gene transfer.

Chemical Methods:
DEAE-dextran (Diethylaminoethyl-dextran)- It is polycationic compound and is derived from dextran (a polymer of carbohydrate). DEAE-dextran is capable of binding to the anionic phosphodiester backbone of the deoxyribo nucleic acid (DNA) due to its positive charge. The resulting complex retains an overall cationic charge and is capable of binding to the surfaces of negatively charged cell membrane. Consequently, the complex is then taken up by the cell most probably by the process of endocytosis. The advantages of the DEAE-dextran transfection technique include its easiness, reproducibility, and lower cost.

Calcium Phosphate - Co-precipitation of Calcium phosphate is one of the most famous and broadly utilized techniques for transfection of DNA from the time when it was initially introduced by Van Der Ebin and Graham during early years of 1970’s. This method involves mixing the nucleic acid (DNA) with calcium chloride, and then cautiously adding this mixture to a saline solution with phosphate buffer accompanied by incubating the mixture at room temperature. This produces a precipitate with DNA, which is then spread onto cultured cells. The precipitate is then taken up by the cells by means of phagocytosis or endocytosis. The chief advantages of the calcium phosphate technique are that it is simple, applicable to a wide range of cell types and can be accomplished at lower cost.
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RE: Transformation, Transduction and Transfection –Gene transfer methods - by adimed - 09-12-2013, 12:49 PM
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