genetic engineering

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Principles of genetic engineering: 

Principles of genetic engineering L Mathias

What is genetic engineering: 

What is genetic engineering Genetic engineering, also known as recombinant DNA technology , means altering the genes in a living organism to produce a Genetically Modified Organism (GMO) with a new genotype. Various kinds of genetic modification are possible: inserting a foreign gene from one species into another, forming a transgenic organism ; altering an existing gene so that its product is changed; or changing gene expression so that it is translated more often or not at all.

Basic steps in genetic engineering: 

Basic steps in genetic engineering Isolate the gene Insert it in a host using a vector Produce as many copies of the host as possible Separate and purify the product of the gene

Step 1: Isolating the gene: 

Step 1: Isolating the gene

Step 1: Alternative method (using reverse transcriptase): 

Step 1: Alternative method (using reverse transcriptase) Reverse transcriptase mRNA converted into cDNA Complementary strand produced using DNA polymerase Advantage – more mRNA in cell than DNA

Step 2: Inserting gene into vector: 

Step 2: Inserting gene into vector Vector – molecule of DNA which is used to carry a foreign gene into a host cell

Step 3: inserting vector into host: 

Step 3: inserting vector into host See worksheet

Replica plating: 

Replica plating

Step 4: Multiplication of the host cells by cloning: 

Step 4: Multiplication of the host cells by cloning Large scale fermenters by cloning All genetically identical because of asexual reproduction

Step 5: Extraction of desired gene product: 

Step 5: Extraction of desired gene product