What is the process of bacterial transformation? Why is it difficult to get fore
ID: 60534 • Letter: W
Question
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?
Describe how restriction enzymes are used in the process of making recombinant DNA molecules. Why is ligase needed to make recombinant DNA molecules?
How are “selectable markers” such as antibiotic resistance genes used in cloning and specifically bacterial transformations?
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.”
Explanation / Answer
1. What is the process of bacterial transformation?
A - Transfer of genetic material [exogenous] into the bacterial cell either by uptake from the external medium or with the aid of phage.
2. How are bacterial cells made “competent” to receive DNA?
A- By treating them with CaCl2 makes the cell wall permeable to DNA as pores are created in the cell wall, from where DNA enters the cells force fully upon heat shock treatment.
3. Why is ligase needed to make recombinant DNA molecules?
A - Ligase helps to bind the fragments together as a single unit for forming recombinant DNA. It forms the phosphodiester bond between the nucleotides.
4. How are “selectable markers” such as antibiotic resistance genes used in cloning and specifically bacterial transformations?
A - This is because if the plasmid is integrated into the bacterial chromosome, it will disrupt the resistance gene and cells will not show resistance, and if not then they will be resistant to the antibiotic.
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