Question

1. List each of the seven steps involved in rDNA technology, from obtaining large amounts of this gene to confirming your pure protein X gene from an E. coli and describe one of them briefly (2 pts).

Answer 1

Recombinant DNA technology can be described as a technology in which DNA of two different organisms is merged and put into a host. This technology allows the production of genes which are beneficial to humans.

The steps of the rDNA technology are:

1) Isolating of DNA fro a cell

2) Using Restriction Enzymes to cut the DNA

3) Isolation of the desired DNA

4) Amplification of the desired gene

5) Ligating the DNA fragment into a suitable vector

6) Transfer of the foreign DNA into an host organism

7) Getting the gene product

One of the process to describe will be:

Amplification of the desired gene: The most common method of amplifying a gene in vitro is by the technique of PCR.

There are three basic steps of PCR:

Denaturation:

In this step, the two strands of the DNA are separated by heat. Usually the temperature is set to 92-95 degrees for 15 sec Celsius for this process.

Annealing:

In this step, primers are used to anneal the DNA strands. This step is carried out at lower temperatures mainly in between 60-65 degrees Celsius.

Extension: The Taq polymerase enzyme adds nucleotides to the primers complementary to the DNA strands.