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Protein expression problem

Protein expression problem - Molecular Cloning Forum

Protein expression problem - DNA cloning forum. Discuss the cloning of plasmids, vectors, DNA, RNA, cDNA, proteins, and other molecules for expression, sub-cloning, and other methods. Also discuss digestion, ligation, transformation, plasmid preparation, cDNA library screening, and competent cells here.


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Old 01-26-2009, 07:43 AM
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Default Protein expression problem



Hi,
I have been trying to express protein in HeLa cell with pcDNA3.1 mammalian expression vector. I had to use his-tag at c terminal of my target protein since my protein cant be detected with suitable antibody. The problem is I found very low expression of my protein. I checked also with Ds-Red system and saw expression under microscope but it is low. Can anybody suggest me some strong mammalian expression system to increase my protein expression?? I was thinking of pMEX-HA expression system..any idea? Thanks in advance.

Zahed
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Old 02-14-2009, 03:00 AM
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Default Re: Protein expression problem

Hi Zahed,
have you tried different cell lines? Also try to use a yeast cell line expression system that secretes your protein into the media - much easier to collect and purify.
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