Recombinant DNA technology was primarily discovered to help researchers determin
ID: 60669 • Letter: R
Question
Recombinant DNA technology was primarily discovered to help researchers determine the function of genes. Why was it difficult for researchers study individual genes before this technology existed?
Why are bacteria used to clone genes?
Describe how restriction enzymes are used in the process of making recombinant DNA molecules. Why is ligase needed to make recombinant DNA molecules?
What are some of the challenges in using recombinant DNA technology to make human therapeutics? Use the example of insulin production described on the DNA interactive web site and describe 2 major problems experienced when producing insulin in bacteria.
What were some of the ethical issues surrounding the use of recombinant DNA molecules that confronted scientists at the time and do they still exist? This discussion can be found again in the DNA interactive site and the module “putting it together.”
What is the process of bacterial transformation? Why is it difficult to get foreign DNA into bacterial cells? How are bacterial cells made “competent” to receive DNA?
Explanation / Answer
It was difficult to study individual genes because there were no systems to isolate genes and study their function in another conducive system.
Bacteria are used to clone genes because they help in replicating the vector construct na dmaking a laege number of copies of such a construct containing an insert of a gene of interest.
Restriction enzymes cleave DNA at specific sites in either sticky or blunt configuration. Ligase enzyme joins two such pieces of cut DNA.
The biggest problem of using recombinant DNA technology in bacteria is due to the fact that the regulation systems of a prokaryote and a eukaryotic system are different. Thus, such an insulin produced would not fold properly and lack the major posttranslational modifications required for a fully active insulin.
The process of uptake of genetic material (DNA) from external sources by crossing the plasma membrane is called a transformation. It is difficult to get foreign DNA because DNA is negatively charged and the positively charged amino acids on bacterial cell walls do not allow its entry. Cells are made competent by several methods such as the calcium chloride method to improve the adherence of DNA on the cell membrane and finally introduce it inside the bacteria.
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