Y2H所需材料：.doc

Y2H所需材料：　 PJ69-4A PJ69-4α or AH109 Y187　 　 pGBKT7 DNA-BD Vector (bait)pGADT7 AD Vector (prey)pGBKT7-53 Control VectorpGBKT7-Lam Control Vector　　 pGADT7-T Control VectorpCL1 Control Vector　　 3 DNA-BD AD Sequencing PrimersHerring testes carrier DNA　　 Peptone，Yeast extract，Dextrose(glucose)； SD base； DO Supplement；　　　　　　Yeast Phenotypes TABLE I. LIST OF ABBREVIATIONS 　 Ade–, His–, Leu–, or Trp–　　　 Requires adenine (Ade), histidine (His), leucine (Leu), or tryptophan (Trp) in the medium to grow; is auxotrophic for at least one of these specific nutrients.　　　 Ade+　　　 Expresses the ADE2 reporter gene; i.e., does not require Ade in the medium to grow.　　　 His+　　 Expresses the HIS3 reporter gene; i.e., does not require His in the medium to grow.　 LacZ+　　　 Expresses the lacZ reporter gene; i.e., is positive for b-galactosidase activity. 　 Mel1+　　 Expresses the MEL1 reporter gene; i.e., is positive for a-galactosidase activity. 　 Miscellaneous 　 Ade2p　　 Protein encoded by the yeast ADE2 gene. 　 3-AT　 3-amino-1,2,4-triazole; a competitive inhibitor of the His3 protein. 　　　 CHX　 Cycloheximide 　　 DO　　 Dropout (supplement or solution); a mixture of specific amino acids and nucleosides used to supplement SD base to make SD medium; DO solutions are missing one or more of the nutrients required by untransformed yeast to grow on SD medium. 　　　 His3p　　 Protein encoded by the yeast HIS3gene. 　 SD medium　 Minimal Synthetic Dropout medium; comprised of a nitrogen base, a carbon source (glucose or galactose), and a DO supplement.　　　 YPH　　　 Yeast Protocols Handbook 　　　X) X-a-gal（with DMF） TE buffer DMSO PEG/LiAc (10X)　　　Strain　 SD/–Ade　　 SD/–Met　　 SD/–His　　 SD/–Ura 　　　 YPDA　　 YPD/CHX　　 SD/–Leu　　　 SD/–Trp　 PJ69-4A　　 –　 +　　　–　　　 +　　 +　　 –　　 –　　　 –　　　 PJ69-4α　 –　　　 –　　　–　　 +　　 +　 –　　 –　 –　　　 　　　　　　 Fusion　　 EpitopeYeast selection　　 Bacterial selection　 Cloning vectors　 　　 　　 　 　　　 pGBKT7　 DNA/bait　 c-Myc　　　 TRP1　　　 kanamycin　 pGADT7　　　 AD/library　 H