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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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07-22-2008, 02:44 AM
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 deletion mutants Hello, I have made a recombinant protein using PQE9 vector in Ecoli. currently, I'm trying to make some deletion mutants. I have done making the deletion mutants DNA using PCR; and ligated them into the vector (sticky ends). but the proteins are not expressed. Please, anyone has any comment!        |
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07-22-2008, 06:16 AM
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 Re: deletion mutants Hello Goodluckewan, welcome to the forum. have you sequenced your vectors in both directions? Also have you made sure your protein is in sense-coding frame with the transcriptional start site (ie have you checked the triplets after the start)? Also how are you detecting your proteins? Maybe detection is the problem. cheers  |
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07-22-2008, 10:11 PM
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| Re: deletion mutants Thank you, and I am going to check sequences. |
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