Question

1. During a transient transfection assay?

a. DNA is introduced into a eukaryotic cell and remains extrachromosomal

b. DNA is introduced into a eukaryotic cell and is stably intergrated into a chromosome.

c. DNA is introduced inot bacterial cells for amplification.

d. DNA is taken up and package in viral capsids.

2. You want to study regulatory that controls expression of a gene whose protein production is difficult to measure quantitatively. What method should you use to analyze gene expression?

a. Western blot with an antibody specific for the person of interest

b. reporter gene assay

c. DNAse I footprinting

d. coimmunopreciptitation assay

Answer 1

1. During transient transfection assay the (a) DNA is introduced in the Eukaryotic cell and it remains extrachromosomal

In Transfection process, the foreign genetic material is introduced into a Eukaryotic cell to study the altered gene expression. This is accomplished physically and chemically. In Transient transfection assay the foreign gene donot integrate with genome and will not replicate and will be lost through cell division. Therefore transient transfection is used to study effects of short term expression if gene or gene products.

2.  To study regulatory that controls expression of a gene whose protein production is difficult to measure quantitatively, method used to analyze gene expression is (b) Reporter gene assay.

The reporter gene is fused with the gene of interest and cloned into an expression vector. The expression vector with the chimeric sequence is then transferred into suitable cells and the expression of the gene in the cell is measured by the activity of the reporter protein.

Western blot is used to identify proteins qualitatively. DNAse I footprinting is used to determine DNA protein interaction. Coimmunopreciptitation assay technique to identify relevant protein–protein interactions by using protein-specific antibodies and to isolate a protein.