baculovirus and PCR

Steve Rodems smrodems at students.wisc.edu
Sun Mar 27 23:26:18 EST 1994


In article <2n5l7v$k3n at server.st.usm.edu>, sywang at whale.st.usm.edu (Shiao
Y. Wang) wrote:

> 
> I have a pair of primers that I use to detect baculovirus infected shrimp.
> The problem is that the PCR reaction doesn't always work. Since
> baculovirus causes that production of occlusion bodies (protein crystals
> containing virions), one possible reason is that simply boiling the
> infected tissue in water does not release the DNA for amplification. I
> would be grateful for information on how you extract DNA from tissues for PCR.

Production of occlusion particles is one of two forms of baculovirus
progeny.  The other form is the budded virus particle which is responsible
for cell-to-cell transmission (occlusion particles are responsible for
organism-to-organism transmission).  To make a long story short, there
should be plenty of budded viral particles within the tissues and released
from cells that you can PCR amplify.  Whether or not boiling tissue is
enough to extract them, I don't know.  I have only used tissue culture and
I conveniently get budded virus in cell culture medium.  I use detergent
(Tween-20 and Triton X100) and Proteinase K to remove the protein coat of
the virus.

-- 
Steve "Some day I will get the hell out of Wisconsin" Rodems

"Then I am here for the Lee family renioun ... shur-wajo-shur"



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