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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

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【作者】胡丽娟；鲁芳；汪宇辉；刘德松；刘彦友；肖静；朱彬；郭慧玲；万朝敏；王正荣；

【Author】 HU Li-juan~(1),LU Fang~(2),WANG Yu-hui~(1),LIU De-Song~(3),LIU Yan-You~(1),XIAO Jing~(1),ZHU Bin~(1),GUO Hui-Ling~(1),WAN Chao-Min~(4),WANG heng-rong~(1)(1.Dept of Biomedical Engineering,School of Basic Medicine and Forensic,West China Medical Center,Sichuan University,Chengdu 610041,China;2.Molecular Biology and Genetics Center,Sichuan Academy of Medical Science & Sichuan Provincial Hospital,Chengdu 610072,China;3.Dept of Parasiotology and Lab of Morphology,School of Basic Medicine and Forensic,West China Medical Center,Sichuan University,Chengdu 610041,China;4.Section of clinic epidemiology,Second University Hospital,West China Medical Center,Sichuan University,Chengdu 610041,China)

【机构】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；四川省医学科学院.四川省人民医院检验科分子遗传中心；四川大学华西医学中心基础与法医学院形态学实验室；四川大学华西妇产儿童医院儿科；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；成都610072；成都610041；

【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.更多还原

【Abstract】 The PAS domain of hPer1 cDNA was amplified by PCR.The amplified fragment of hPer1 cDNA was ligased with the vector pGBKT7 to construct recombinant bait plasmid pGBKT7-hPer1_PAS.The cDNA library 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-hPer1_PAS was constructed correctly.The transformation rate was about 1.4×10⁶/3μg pGADT7-Rec.fourty six colonies were selected by the yeast two-hybrid system.更多还原

【关键词】近日节律PER1；蛋白相互作用；酵母双杂交；
【Key words】 PER1； recombinant bait plasmid； cDNA library； yeast two-hybrid；

【基金】由国家自然科学基金资助(304706233007028830470684)

【文献出处】四川大学学报(自然科学版) ,Journal of Sichuan University(Natural Science Edition) , 编辑部邮箱 ,2006年05期

【分类号】Q78
【被引频次】3
【下载频次】119

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