Question on reverse transcriptase(s)
axa12 at po.CWRU.Edu
Sun Aug 4 11:04:35 EST 1991
I have a question/survey about reverse transcriptases.
There are two enzymes that are commercially available, the MuLV
and AMV reverse transcriptases (RTs).
The differences between the two are as follows:
a) 2 subunits 1 subunit
b) active 45 degrees active 42 degrees
c) optimum temp = 42 degrees optimum temp = 37 degrees
d) contains intrinsic no endonuclease activity
e) No RNase H mutant is RNase mutant is
commercially available sold commercially
f) Purified from virions Bacterially produced
g) The AMV enzyme is supposed to have greater processivity than
the MuLV enzyme
As part of my Ph.D, I am in the process of cloning the two subunits of
the avian enzyme (AMV RT). I am also making some mutations in the
enzyme, including one that knocks out the intrinsic endonuclease.
Although there is no reason for me to do this, I could also fairly
easily make one that knocks out the RNase H activity, without
affecting processivity. If I made this mutation, I would be interested
in making it freely available.
My question is:
"Given your choice of the following 4 enzymes for a cDNA synthesis,
or primer extension experiment, which would you use?"
a) Native MuLV RT (no endonuclease, possesses RNase H)
b) Altered MuLV RT (no endonuclease, RNase H knocked out)
c) Native AMV RT (intrinsic endo, possesses RNase H)
d) Altered AMV RT (no endonuclease, RNase H knocked out)
Please mail your response and comments to axa12 at po.cwru.edu
Graduate Student in Biochemistry
Department of Biochemistry
CWRU Medical School
Cleveland, OH 44106-4935
My views (and deeds) are my own, and Case Western Reserve University
is not responsible for any of them.
axa12 at po.cwru.edu
aiyar at cwbio.bioc.cwru.edu
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