Questions on Baculovirus system

Huseyin Kucuktas kucukhu at mail.auburn.edu
Fri Jan 6 23:56:20 EST 1995


	Hello,

	I am trying to express a protein which is supposed to be a 
product of a 5406 bp DNA fragment. However the actual ORF is small and 
the gene reads on the complementary strand.
	 I HAVE SOME QUESTIONS!

	1-  Maximum how many extra bases I can have before the ATG 
codon of my "gene"? 
	2-  It appears that I do not have a lot of desirable restriction 
sites around the beginning of the ORF, how am I going to get rid of the 
extra bases?
	3-  I am planning to use polyhedrin promoter based vectors. The 
kits that the companies sell are very expensive (almost $500 for 5 
transfections). Which one is the best choice? Is occ based selection too 
difficult?


	Thanks for the help! 



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