Clontech for baculovirus? NOT! NUNCA! NIEN!

Gregg Silk greggsilk at aol.com
Mon Jun 15 23:35:44 EST 1998


Sorry if there was any confusion in the previous posting. I had not expected to
get this involved, so I just went downstairs for the Bac-to-Bac manual.  

Clontech makes BacPac.  

Life Technologies (GibcoBRL) makes Bac-to-Bac which uses the pFastBac (FB)
vector as the donor plasmid. I did this 18 months ago, and even then  all the
"bac-this" and "bac-that" got confusing at times. 
Since I have this stuff in front of me..... 
Bac-to-Bac (FastBac) (FB) (LifeTech) (GibcoBRL)  has:
High efficiency recombination in a special Ecoli host strain.
Strong selectable ANTBIOTIC markers for recombinant virus,  not wussy
complementation.
Screen and grow bacmid in Ecoli before transfection.
No nonrecombinant virus background, avoids  marathons of plaque purification.
Efficient transfections using standard reagents. 
MOST important, the limiting reagent is not obscenely expensive linear virus
(like certain other companies). The imiting reagent is  the host Ecoli cells
which  were $15 a tube and work very well.

Is expression cloning simple? No, it's still a big committment and it takes
very savvy planning, but there is a reasonable chance of success. 
Your mileage may vary  ;)

Gregg



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