(none)

Katja Aviszus aviszusk at NJC.ORG
Wed Jun 24 19:00:49 EST 1998


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Hi netters,
I'm designing a phage expression vector for Ab genes.  The 3' of the heavy =
chain will be joined to a c-myc tag which is then joined to protein 3 from =
filamentous phage.  The constructs that I have found separate the 3' of the=
 heavy chain from the c-myc tag with a NotI site (ala ala ala).  Between =
the c-myc and the protein 3 is (gly gly gly gly ser).  Because of the way =
I have to build my vector, I have to incorporate other amino acids but I =
am afraid they will interact with other sequences.  Between c-myc and =
protein 3 I am going to have to add 2 amino acids to give (thr ser gly gly =
gly gly ser).  Between the 3' end of the heavy chain and c-myc I may have =
to add 2 amino acids as well to give (thr ser ala ala ala).  I can avoid =
adding the thr ser in this one by doing site directed mutagenesis at 2 =
locations in the vector.  Is it worth doing the site-directed mutagenesis =
to avoid the 2 extra amino acids?  Do you think the (thr ser) will cause =
any problem interactions in either situation?

Thanks ahead of time for your help.  Any input would be great.  my address =
is

shapirog at york.uchsc.edu


Thanks,
Gary Shapiro

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<HTML><HEAD></HEAD><BODY><FONT FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#000000">=
Hi netters,<BR>
I'm =
designing a phage expression vector for =
Ab genes.  The 3' of the heavy chain will =
be joined to a c-myc tag which is then joined =
to protein 3 from filamentous phage.  The =
constructs that I have found separate the =
3' of the heavy chain from the c-myc tag =
with a NotI site (ala ala ala).  Between =
the c-myc and the protein 3 is (gly gly =
gly gly ser).  Because of the way I have =
to build my vector, I have to incorporate =
other amino acids but I am afraid they will =
interact with other sequences.  Between =
c-myc and protein 3 I am going to have to =
add 2 amino acids to give (thr ser gly gly =
gly gly ser).  Between the 3' end of the =
heavy chain and c-myc I may have to add =
2 amino acids as well to give (thr ser ala =
ala ala).  I can avoid adding the thr ser =
in this one by doing site directed mutagenesis =
at 2 locations in the vector.  Is it worth =
doing the site-directed mutagenesis to avoid =
the 2 extra amino acids?  Do you think the =
(thr ser) will cause any problem interactions =
in either situation?<BR>
<BR>
Thanks ahead of =
time for your help.  Any input would be =
great.  my address is<BR>
<BR>
</FONT><FONT FACE=3D"Monaco" =
SIZE=3D1 COLOR=3D"#0000FF"><U>shapirog at york.uchsc.edu</U></FONT><FONT =
FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#000000"><BR>
<BR>
<BR>
Thanks,<BR>
Gary =
Shapiro<BR>
</FONT></BODY></HTML>
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