ISSN:
1573-9368
Keywords:
40.6 transgenic mouse
;
MutaMouse
;
mutant selection
;
λgt10lacZ phage
;
galE
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract The determination of thelacZ mutant frequency in λgt10lacZ phage vectors isolated from the transgenic mouse strain 40.6 (MutaMouse), requires the screening of large numbers of phages on β-galactosidase activity. Existing methods rely on distinguishing a few white plaques on X-gal containing plates amongst a multide of blue ones which is both time-consuming and expensive. The new screening method described here employs the galactose sensitiveEscherichia coli C lacZ recA galE strain into which a multicopy plasmid has been introduced, which results in over-expression of thegalK andgalT genes. In the presence of phenyl-β-d-galactopyranoside, a substrate for β-galactosidase, this leads to the suppression of λlacZ + phage propagation without affecting the ability of λlacZ − phages to form plaques. With this method it is possible to screen 1.5×106 phages on a single 9-cm Petri dish. Furthermore, the need for blue/white screening has been eliminated.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01976029
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