Packaging cell-lines for virus

Greg Tobin tobin at fcs260c.ncifcrf.gov
Fri Dec 2 13:30:04 EST 1994


In article <9411017863.AA786302947 at internetmail.pr.cyanamid.com> Bart_Corsaro_at_USLRMG01 at INTERNETMAIL.PR.CYANAMID.COM writes:
>
>----------------------------Original message----------------------------
>Hi, Netters!
>   I am trying to learn how to make virus-like particles so that I
>will ultimately be able to determine the N-terminal sequence of a protein
>not made in abundance by the native virion.  (This method was vaguely suggested
>to me by the study section which reviewed my grant, and I don't know a lot
>about it.)  I have been reading a little here and there about the "psi" line
>for packaging virus, and I am searching for a comprehensive review.  Does
>anybody know of one?  Thanks a lot.  Post or Email reply, please.
>LKM100F at ODUVM.cc.odu.edu
>-------------------------------------------------------------------------------
>Laura
>
>The baculovirus expression vector system has been used to produce virus-like 
>particles for several viruses including Rotavirus, Human Papilloma Virus and 
>Calicivirus. Unfortunately, this system requires you to be able to clone the 
>capsid genes into a baculovirus transfer vector which you may or may not be 
>able to do.
>

Baculovirus expression vectors have been used to express almost every
known retrovirus Gag precursor.  Most precursors self-assemble and
bud like virus particles.  However, expression of the pol genes 
with the gag genes results in cytoplasmic viral protease activity
that digests the Gag precursor before it can form a particle.  This
may not be in the literature.  In addition, the psi 
sequence may not function well in insect cells as none of the 
particles that I have made package virus-specific RNA - which is 
an added feature for their initial design as parts of putative vaccines
six years ago.



-- 
Greg Tobin, Ph.D.                        tobin at lcms-1.ncifcrf.gov
LCMS 
PO Box B
Frederick, MD 21702



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