DNA Construction: HIV-1 Tat Gene
Posted 19 March 2012 - 03:03 AM
I am interested in getting the longer variant (101 a.a. residues) of the protein encoded, ie. one of which the translation stops at the second stop codon. And a search of commercially available plasmid vectors carrying this gene include both stop codons. and in these plasmid vectors, the 2 exons are separated, unlike in my construct.
So my question is: Should I include the first stop codon in my construct, or should I remove it as there is no way I can tell if my full construct, which contains many other non-HIV elements as well, is 'native' enough for the translation machinery to ignore the first stop and proceed until the second one.
Thanks in advance.
Posted 19 March 2012 - 03:23 AM
Posted 19 March 2012 - 03:26 AM
But it is found in, for example, HXB2 and NL4-3, no?
Posted 19 March 2012 - 04:18 AM
Posted 19 March 2012 - 05:47 AM
Thanks! Do you have any experience in constructing the Tat gene? Do you change the premature Stop codon to Serine (TAG to TCG), since Serine is the amino acid residue present in SF2 in place of the Stop codon in other strains, or to other amino acid residues?
Posted 19 March 2012 - 07:31 AM
Posted 19 March 2012 - 09:20 AM
Hey man, thanks! But this sequence codes for more than 101 a.a. residues. Strange. Anyway, I have aligned the Tat gene sequence from 8 different HIV-1 strains. Of these 8, 4 have a premature stop codon, 3 have a serine in place of the premature stop codon and the last one has a glutamine.
So I will construct a Tat gene with a serine residue. Thanks a lot!! You've been a star!
Posted 19 March 2012 - 10:12 AM
Posted 19 March 2012 - 10:14 AM
Posted 19 March 2012 - 10:22 AM