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| Molecular Cloning Forum 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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#1
01-10-2010, 01:20 PM
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 His-tag fusion protein in pET-28a(+) Hi. I am student only getting familiar with gene cloning. I want to insert a gene in this vector (pET-28a(+), in order to get a his-tag. My question is: do you have to worry about the stop codon of the insert itself, because the vector already has a stop codon after the His-tag. start--Histag---(start - Coding sequence -stop)----Histag--stop. So, there are two stop codons, how do i know that translation does not stop before the his-tag is completely incorporated? Thanks, fragnatic  |
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#2
01-11-2010, 07:13 PM
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| Re: His-tag fusion protein in pET-28a(+) Those are not the same His tag. There is an N terminal, and optional C terminal tag if you do not incorporate a stop codon. |
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| Tags |
| fusion , fusionprotein , his-tag , histag , pet28a , protein , stop codon , vector |
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