Name 3 reasons a scientist would transform bacteria Describe

Name 3 reasons a scientist would transform bacteria.

Describe 3 methods to determine if your transformation was successful.

What might have happened to interfere with a successful transformation?

Solution

Answer:

3 reasons a scientist would transform bacteria are:

(1) For the overexpression of a particular protein in an expression host (such as BL21-DE3 cells)

(2) For cloning a gene of interest and corresponding isolation of plasmids containing the gene for further use, such as sequencing, subcloning, etc.

(3) Preparation of cDNA libraries.

3 methods to determine if the transformation was successful are:

(1) Appearance of resistant colonies (bacteria which got transformed with the plasmid containing the antibiotic resistant gene) on antibiotic plates.

(2) Blue-White colony selection with X-gal substrates. The white colonies are the transformed ones.

(3) Isolating the plasmid from the colonies obtained and testing for the presence of the plasmid containing the insert through restriction digestion.

A number of factors may affect the transformation efficiency:

(1) Competency of the bacterial cells being used for transformation- Less competent cells shall give poor results.

(2) Methods of transformation- Electroporation is more efficient than chemical transformation.

(3) Forms of DNA- Supercoiled DNA gives better results in a transformation experiment.