question about protein translation

sdyang sdyang68 at yahoo.com.cn
Sat May 8 11:00:00 EST 2004


Hello, Everyone:

 

  If you have used a SMART TM cDNA Library Construction Kit produced
by Clontech company to construct cDNA library, you may find that there
was a slight change between kits produced three years ago and those
produced at present. The difference occured in the nucleotide sequence
of SMART III primer. There was a ATG codon (in the Sfi I recognise
site) in the SMART III primer of the old kits, while this ATG codon
have been changed in the SMART III primer of the new kits. Therefore,
the cDNA constructed using old kits will be brought an additional ATG
codon at the 5' end. if you insert cDNA into vectors at Sfi I site,
the ATG codon will be remained.

  I want to know the reason why the Clontech make this change since I
have use the old kits to constructed two cDNA librarys. Furthermore, I
have use one of this cDNA library to transform yeast in order to
isolation genes by yeast function complementation. Because all the
cDNAs were inserted into E. coli - yeast shuttle expression vector at
Sfi I site.

  I really wander the genes can be translated from the ATG codon
provided by the kits, not from their native ATG. If it is the truth,
the consequence will be terrorful since I must withdraw my whole
results of yeast function complementation. I eagerly want you tell me
what I worried can be occur or not. I also hope you can tell me
whether there are several translation products or not for a gene have
several ATG codons. thank you.

your sincerely 
Shude Yang

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