Alternatives to Isogenic Mutants

[bwschn01]

I've been tasked to come up with alternatives to generating an isogenic mutant bacteria for studying gene function and phenotype. I've come up with the conditional mutants. I'm hoping to find at least one more alternative. Can anyone direct me to a protocol or a paper?

 

Thanks,

bwschn01

[iRNAblogger]

Hi bwschn01. Would you mind giving a little more details about the goal of your experiments? Just a few basic suggestions (that I'm sure you have probably considered) come to mind, but if you gave a little more information about exactly what kind of experiments you would like to run, then I could try to give more specific suggestions.

 

DISCLAIMER: I am not a microbiologist, but I have done some bacterial genetics; So I'm not sure how helpful I will be.

 

For phenotyping, you might consider constructing an ectopically regulated strain (for the gene in question). That way you could control exactly when it is expressed without actually mutating the protein coding sequence. You could also try replacing it with homologous genes from related species to see if the phenotype is the same. (I'm not sure which species you are working with, so these might not be possible)

 

For functional analysis you could try a pulldown to see what interacting proteins come with it. Other than that you might consider running HHPRED on the gene's protein sequence to see if anyone has studied a related protein (bioinformatic approach).

Edited October 17, 2013 by iRNAblogger