HELP! Problems with the expression of HSP 70.

sl16s at cc.usu.edu sl16s at cc.usu.edu
Fri Nov 18 10:39:35 EST 1994


I am trying to express a fungal HSP70. I have tried pUR278 vector first.  I
cloned my insert in the three different reading frames. but did not see any
expression. I changed the vector to pET 21 and subcloned into the three
different RFs again. but still not able to see any expression.  I know the
first 300 and last 300 base pairs of the insert sequence. There is a start
codon and a poly A tail.
        My questions are:
        1. Could there be  a stop codon introduced into my insert and the read
        ing is stopped?.
	2.  Is thereany possibility of not detecting protein eventhough it is 	
produced?.         
(I did not use PCR to clone my insert, it is a cDNA insert)
Thanks in advance.
					Sincierly
					Sujatha paul.




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