利用酵母双杂交研究铜绿微囊藻生物钟蛋白KaiB的自身相互作用  

Study on self-interaction of clock protein KaiB of Microcystis aeruginosa by yeast two-hybrid

作　　者：郑锦乾[1] 徐虹[1] 章军[1] 王靖[1] 朱斌琳[1] 

机构地区：[1]厦门大学生命科学学院细胞生物学与肿瘤细胞工程教育部重点实验室,福建厦门361005

出　　处：《海洋科学》2006年第8期41-45,共5页Marine Sciences

基　　金：国家自然科学基金资助项目(40306024)

摘　　要：从铜绿微囊藻(Microcystis aeruginosa)PCC7820中克隆了生物钟基因kaiB,并将其克隆入酵母双杂交系统的两个质粒pGADT7和pGBKT7中。经测序验证后,将重组质粒pGADT7-kaiB和pGBKT7-kaiB转化酵母菌AH109进行自激活性和自身相互作用的检测和验证。结果表明,铜绿微囊藻生物钟蛋白KaiB无自激活性,KaiB蛋白自身能发生相互作用。因此,可用KaiB蛋白为诱饵利用酵母双杂交系统筛选文库钓取与KaiB发生相互作用的蛋白。Clock gene kaiB of Microcystis aeruginosa was amplified and cloned into pGBKT7 and pGADT7 respectively. After being verified by sequencing, the two recombined plasmids pGADT7-kaiB and pGBT7-kaiB were transformed into yeast strain AH109 by polyethylene glycol/lithium acetate method and its expression products were assayed whether it can affect the growth of yeast cells and activate the reporter （β-galactosidase） genes. The result shows that KaiB protein was not toxic to AH109 and could not activate the reporter gene. KaiB protein could play self-interaction. Yeast two-hybrid GAL4 system could be applied to fish KaiB interacting protein.

关 键 词：铜绿微囊藻(Microcystis aeruginosa) 生物钟蛋白KaiB 酵母双杂交 自身相互作用 

分 类 号：Q78[生物学—分子生物学]