酵母双杂交筛选血液中与PERIOD1相互作用的新蛋白  

Selection of the Novel Proteins Interacting with hPER1 in Blood

作　　者：胡丽娟[1] 鲁芳[2] 汪宇辉[1] 刘德松[3] 刘彦友[1] 肖静[1] 朱彬[1] 郭慧玲[1] 万朝敏[4] 王正荣[1] 

机构地区：[1]四川大学华西医学中心基础与法医学院生物医学工程研究室,成都610041 [2]四川省医学科学院.四川省人民医院检验科分子遗传中心,成都610072 [3]四川大学华西医学中心基础与法医学院形态学实验室,成都610041 [4]四川大学华西妇产儿童医院儿科,成都610041

出　　处：《四川大学学报（自然科学版）》2006年第5期1127-1131,共5页Journal of Sichuan University(Natural Science Edition)

基　　金：由国家自然科学基金资助(304706233007028830470684)

摘　　要：采用酵母双杂交方法,以人PER1的PAS结构域为诱饵,在人血液cDNA文库中筛选能与之相互作用的蛋白.经酶切和核苷酸序列测定,证实重组诱饵载体hper1PAS/pGBKT7构建成功.血cDNA文库转化效率为1.4×106/3μg pGADT7-Rec.酵母双杂交文库营养缺陷筛选得到114个阳性克隆,β-半乳糖苷酶检测报告基因获得46个蓝色克隆.The PAS domain of hPerl cDNA was amplified by PCR. The amplified fragment of hPerl cDNA was ligased with the vector pGBKT7 to construct recombinant bait plasmid pGBKT7-hPer1pAs. The cDNA li- brary of human blood was constructed. The recombinant plasmid and the cDNA library were transfected into yeast strain AH109 by LiAc Method. The yeast two-hybrid system was screened by SD/-Ade/-His/-Leu/- Trp. Results Digested with endonuclease and sequenced, the recombinant plasmid of pGBKT7-hPer1PAS was constructed correctly. The transformation rate was about 1.4 ×10^6/3μg pGADT-Rec, fourty six colonies were selected by the yeast two-hybrid system.

关 键 词：近日节律PER1 蛋白相互作用 酵母双杂交 

分 类 号：Q51[生物学—生物化学]