E. coli lacZ complementation

mike BENEDIK at UH.EDU
Thu Oct 20 16:56:46 EST 1994




In article: <199410141622.JAA23090 at net.bio.net> 
hwyckoff at ASRR.ARSUSDA.GOV ("HERBERT WYCKOFF") writes:
> 
> 
> I would like to modify an E. coli strain(MC1061) that is much used in
> our lab so that it can be used to perform blue/white color screening
> with alpha-complementing vectors.  I would prefer to do this without
> adding any antibiotic resistance to the strain.  If anyone knows of a          
> reference where this type of construction is described, I would 
> appreciate you sending me the citation. Thanks


MC1061 is a complete deletion of Lac. You have 2 options to make it
into
a blue white strain. 

1) get the phi80 dlac phage used in JM83 or similar strains and make
a lysogen of MC1061 with it.

2) make a ProAB- mutant of MC1061 and then mate in the F'pro-lac
episome
used in JM101 or other such strains.

It would be the easiest to use an antibiotic marked episome because
then
no further strain constructions would be needed.

MICHAEL BENEDIK                                          BENEDIK at UH.EDU
BIOCHEMICAL SCIENCES
UNIVERSITY OF HOUSTON
HOUSTON, TX 77204-5934



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