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 Hemoglobin protein alpha-1 and 2 
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Joined: Fri Oct 08, 2010 6:08 am
Posts: 58
Post Hemoglobin protein alpha-1 and 2
Hi guys,

I was constructing a hemoglobin sequence to put into E.coli. The hemoglobin protein structure is made of four proteins, alpha-1, alpha-2, beta-1, and beta-2. The sequences for alpha-1 and 2 are identical. To produce the tetramer protein, do the two sequences need to be repeated? I think that that is what has to happen so that the two proteins are synthesized to make the protein structure. Please provide comments.

Thanks,

chimeraboy


Mon Jan 17, 2011 11:31 pm
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Joined: Thu Aug 19, 2010 3:18 am
Posts: 26
Post Re: Hemoglobin protein alpha-1 and 2
HBA1 and HBA2 differ in their untranslated regions. However, for producing in E. coli, all you want is the coding region. For this, the two are identical and you only need one of the alpha subunits.

An important caveat, however, in producing hemoglobin in E. coli. Typically the protein is misfolded unless the E. coli is grown on media rich in heme. The heme is necessary to stabilize the folding and formation of the tetramer. Not going to lie, producing a multi-subunit protein in E. coli is not the easiest of tasks.


Mon Jan 24, 2011 10:13 pm
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Joined: Fri Oct 08, 2010 6:08 am
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Post Re: Hemoglobin protein alpha-1 and 2
Can you obtain or make heme media? Because from the little research that I have done, the heme needs to be produced genetically. However, if heme could be in the media, that would be super, as currently, the heme-producing genes are taking up a lot of space on the small plasmid that I am planning to work with.

By the way, do you know the efficacy of the hemoglobin chaperone protein and cytochrome b5 reductase in the expression of the hemoglobin tetramer?


Sun Jan 30, 2011 8:52 pm
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