expression constructs & ORF's

converrl at ucbeh.san.uc.edu converrl at ucbeh.san.uc.edu
Mon Nov 14 15:46:47 EST 1994


     I wanted to know if there were any "rules of thumb" regarding overlapping
reading frames when building expression vectors for use in eukaryotic systems. 
     Normally, when I am building a construct for use in eukaryotic cell
expression, I don't worry about overlapping LacZ reading frames from the vector
(such as pBKCMV) which may exist even after trimming away the LacZ promoter
region. My assumption is that in the eukaryotic expression system, the best
context for a ribosome "start" context will be at the ATG position of the
eukaryotic gene I am attempting to clone. I may, however, be mistaken about
this, and I would appreciate any input.
     Also, in bacterial expression systems, I wondered if anyone had any
suggestions as to the minimum distance that can exist (in bp) between one open
reading frame and another open reading frame 3' of it in order for the 3'
reading frame to be expressed in E.Coli.

                      I would greatly appreciate any input!!
                      Please E-mail responses

                      Richard




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