Archita Ghoshal

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Questions related from Archita Ghoshal

Problem with glutathione agarose column for purification of mammalian protein expressed in E.coli - can anyone help?

I have cloned and expressed a mammalian protein in E.coli BL21 with GST tag. I use glutathione agarose beads (sigma) for affinity purification (not pre-packed column). I use 0.4% sarkosyl to...

11 November 2013 2,227 5 View

Expression of only GST protein in pGEX-4t2 containing gene of interest, instead of GST tagged fusion protein?

I have cloned my gene into pGEX-4t2 and expressed in E.coli BL21 DE3. However, my clones are expressing only GST protein. Sequencing results show that my gene has its correct sequence. Does that...

08 August 2013 6,982 11 View

Cloning a mammalian gene into pET28a bacterial vector seems to give toxicity, this is a mysterious problem.

I have cloned a mammalian gene both into pet 28a and pgex-4t2. My protein is expressed properly in BL21 containing pgex-4t2 (I have checked the functional activity of my protein). However, cloning...

06 June 2013 9,131 11 View

What could be the possible reasons for getting a false positive band after doing PCR using plasmid as template?

Quite a few people I know have faced this problem, all using different plasmids and amplifying different insert genes from plasmids. The false positive band is of exact size as the desired insert...

03 March 2013 5,139 17 View

How to analyze PDBsum results?

I docked 2 proteins and analyzed the docked structures using PDBsum. There is no literature stating any residues involved in the interaction. How do I draw a conclusion from the results I have...

10 October 2012 462 10 View