Clontech AH109說(shuō)明書，AH109說(shuō)明書，Clontech AH109

AH109

his section provides detailed phenotypes of the yeast strains included with MATCHMAKER System 3

and MATCHMAKER GAL4 cDNA and Genomic Libraries. For additional information on the growth and

maintenance of yeast, see the YPH, Chapter III. We also recommend Guthrie & Fink’s Guide to Yeast

Genetics and Molecular Biology （1991） and Heslot & Gailardin’s Molecular Biology and Genetic

Engineering of Yeasts （1992）.

A. Yeast Host Strains

The complete genotypes of AH109, Y187, and CG-1945 are provided in Table II. All strains are

gal4

–

 and gal80

–

; this prevents interference of native regulatory proteins with the regulatory

elements in the two-hybrid system.

1. Use AH109 as the host strain if you plan to screen an AD/library using HIS3, ADE2, and MEL1.

2. System 3 Users Only: Use Y187 as the host strain if you plan to test for an interaction between

two known proteins using the lacZ reporter only. In addition, use Y187 as a mating partner to

verify protein interactions.

3. Library Users Only: Use CG-1945 as the host strain if you plan to separate DNA/bait and

AD/library plasmids by cycloheximide counterselection. Alternatively, use pGBKT7 to construct your bait. This DNA-BD vector contains a kanamycin resistance marker; therefore,

vectors can be separated in E. coli without cycloheximide counterselection.