transgenic questions

Lisa Lynn Cunningham llc4q at galen.med.Virginia.EDU
Wed Sep 7 13:31:18 EST 1994


I'm interested in creating transgenic zebrafish, and since I
have no experience with transgenic animals, I have some
questions.  I have read on this newsgroup and heard from some
other discussions that zebrafish transgenics are mosaic.  It
was suggested here that this may be due to use of non-teleost
promoters.  Has any work been done with teleost promoters? My
promoter is a zebrafish promoter, so perhaps I won't encounter
this problem.  My current questions (and I'm sure there will
be others) involve selection of an appropriate vector.  My
first step will be to test my transfection protocol to see that
I am actually getting the DNA into cells.  So I plan to use a
beta galactosidase or luciferase reporter gene in my vector.
Is there an advantage of one reporter gene over the other? Are
there others that I should consider?  What specific vectors
have been used successfully?  In looking at the
commercially-available expression vectors, I see that most of
them contain an SV40 promoter, but some of them are late SV40
and some are early SV40 -- what does the early or late signify?
What are the relative advantages and disadvantages of the two?
Also, since my promoter is tissue-specific, I would eventually
like to see tissue-specific expression of the reporter gene.
Can this be accomplished by merely placing my promoter between
the SV40 promoter and the reporter gene, or will the SV40
promoter need to be removed from the vector to accomplish
tissue-specific expression?




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