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文献和实验Two-hybrid analysis of genetic
combinations of strains,plasmids,and reporters can be used for mating (Section 3).In one common version (Finley and Brent,1994),the strain expressing the bait (bait strain)is RFY206 (MATa ura3-52 his3Æ200 leu2-3 lys2Æ201 trp1::hisG)transformed with the HIS3
EXAMINING THE FUNCTION OF PROTEINS AND PROTEIN NETWORKS WITH THE YEAST TWO-HYBRID SYSTEM
(see below). 7. Test for lacZ expression. One way to do this is simply to patch individual transformants from step 2 to Gal/Raf ura-his- trp- X-Gal plates (about 1 cm x 1 cm patches) and incubate at 30oC. Yeast with a control LexA-activator fusion should turn
培养基 A. SD/Gal/Raf 3.79g B. 10×DO(-His,-Leu,-Trp,-Ura) 10.0ml C. Agar 2.00g D. ddH2O → 85.0ml 灭菌(115℃,10lb/in2)15min ,冷却至50℃,加入下列成份后铺板 E. 10×BU 10.0ml F. 20mg/ml X-gal 0.4ml 90-100mm)f铺5-6块平板( 12. 选择含有pLexA-X显蓝
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