Abstract
The determination of the lacZ 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 sensitive Escherichia coli C lacZ recA galE strain into which a multicopy plasmid has been introduced, which results in over-expression of the galK and galT 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.
Original language | English (US) |
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Pages (from-to) | 67-69 |
Number of pages | 3 |
Journal | Transgenic Research |
Volume | 3 |
Issue number | 1 |
DOIs | |
State | Published - Jan 1994 |
Externally published | Yes |
Keywords
- 40.6 transgenic mouse
- MutaMouse
- galE
- mutant selection
- λgt10lacZ phage
ASJC Scopus subject areas
- Genetics
- Animal Science and Zoology
- Agronomy and Crop Science
- Biotechnology