Recombinant DNA Technology And Genetic Engineering

Recombinant DNA technology or genetic engineering is the use of in vitro molecular techniques to isolate analyze and manipulate fragments of DNA or genes. In the early 1970s, researchers at Stanford University were able to construct chimeric molecules called recombinant DNA molecules. Shortly thereafter, it became possible to introduce such molecules into living cells where they are replicated to make many identical copies. This achievement ushered in the era of gene cloning. Recombinant DNA technology and gene cloning have been fundamental to our understanding of gene structure and function. Recombinant DNA refers to the creation of new combinations of DNA segments that are not found together in nature. The isolation and manipulation of genes allows for more precise genetic analysis as well as practical applications in medicine, agriculture, and industry.

An overview of recombinant dna technology is as follows: Isolate DNA /purifying DNA Cut with restriction enzymes  Ligate into cloning vector  transform recombinant DNA molecule into host cell  each transformed cell will divide many times to form a colony of millions of cells, each of which carries the recombinant DNA molecule (DNA clone).

There are several applications for genetic engineering in microbiology as well as other fields of biology.It includes invitro mutagenesis,gene synthesis, Expressing eukaryotic genes in bacteria,production of transgenic plants and animals,gene therapy,screening for genetic diseases and forensic analysis.

STEPS IN RECOMBINANT DNA TECHNOLOGY

1) PURIFYING DNA/ISOLATION OF DNA

Purifying DNA and isolating genes is an important step in geneticengine...

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...e for the synthesis of new DNA strands, each randomly terminating due to the incorporation of a chain terminating dideoxynucleotide in 4 different reaction tubes. This produces a population of molecules, each terminating at a different site. Running the products in each tube on a gel allows the determination of where each chain terminating dideoxynucleotide was incorporated. The DNA is visualized because the DNA primer to start the reaction is radioactive or some of the dNTPs are radioactive

This procedure is now automated so that a computer reads the sequence. Instead of using radioactive primers, the primers are labeled with different color fluorescent dye for each reaction.

E. PCR (polymerase chain reaction) – Allows the isolation of a specific segment of DNA from a small DNA (or cell sample) using DNA primers at the ends of the segment of interest