Summerresearch

1、生科04級 881714 何國維Lymphocyte stem cell（Bone marrow）B cell （Bone marrow）T cell（Thymus）Tc（Cytotxic T cell）Th（Helper T cell）Th0Th1IFN-T-betADADBDADBD報導基因BDADBDADBDADADADBDADBDReporter geneBDc. Activationb. Fusion protein bindinga. Fusion protein expressionADBDADBDADBDADBDYeast chromosomeConstructed plasm

2、idT-betIFN-r promoterGAL4ADXbaIIFN-r promoterEcoRIpHISi6786 bpHIS3URA3AmpRBamHI (1447)Eco RI (2)Nco I (2935)SmaI (9)XbaI (43)XhoI (1009)XmaI (7)BstXI (542)Eco RV (2918)KpnI (775)SacI (17)HindIII (457)HindIII (644)HindIII (3635)Pst I (836)Pst I (2712)Pst I (3627)ligationReporter geneT-bet binding dom

3、ainEcoRIPstIpGAD4246659 bpLEU2AmpRGAL4ADBamHI (844)Eco RI (834)Pst I (860)SmaI (841)XmaI (839)Eco RV (1891)KpnI (484)HindIII (410)HindIII (1105)BstXI (767)BstXI (1130)BstXI (1930)Activation domainligationReport geneYeast strain：YM4271（MATa）、Y187（MAT）Cant synthesis histidine、 leucine、uracil by itself

4、。YeastIncubateYeast selection medium-His or -Leu or -UraYeast cant growth on selection mediumpHISi-IFN-Yeast（YM4271）Yeast selection medium-HispGAD424-T-betYeast（Y187）Yeast selection medium-LeuYeast selection medium-HisAppropriate 3-AT3-AT：a inhibitor of the HIS3 gene productYeast（YM4271）pHISi-IFN-Yeast selection medium-His/-LeuYeast（Y187）pGAD424-T-betYeast cant growthYeast（YM4271）pHISi-IFN-Yeast cant growthYeast selection medium-His/-LeuIncubateIncubatepHISi-IFN-pGAD424Appropriate 3-ATYeast selection medium-His/-LeuYeast matingYM4271Y187Yeas