Microbiology 6: (Patel) Study protein function

Question 1

Methods for manipulating protein function in cells?

Answer 1

DNA - Over expression of WT, dominant active, dominant negative gene

mRNA (RNA interference)

Protein (Chemical inhibitors)

Question 2

What is transient transfection?

Answer 2

The process of inducing exogenous DNA (genes) into cells

Question 3

What is required to produce recombinant DNA of a GFP tagged protein

Answer 3

cDNA of Protein X
Plasmid vector carrying GFP gene
Restriction Enzymes
DNA ligase

Question 4

in frame cloning RE questions

Answer 4

do some blud

Question 5

What is Site-Directed mutagenesis? Example relating to phosphorylation?

Answer 5

SDM

-> in vitro technique to selectively change one or more amino acids in a protein

Serine residues can be targeted and changed into a negatively charged molecule (Asp, Glu) to mimic phosphorylated state of serine

Question 6

What is RNAi-mediated gene silencing

Answer 6

This term describes a number of related processes which use 21- to 25-nucleotide RNAs to repress the expression of specific target genes