一种与细胞周期素A1互作的新基因的克隆和表达  

Molecular cloning and expression of a novel cyclin A1 interacting gene foca1

作　　者：张尤历[1] 陈劲频[1] 王文兵[2] 

机构地区：[1]江苏大学附属医院消化内科,江苏镇江212001 [2]江苏大学生命科学研究院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2006年第6期465-468,T0002,共5页Journal of Jiangsu University：Medicine Edition

基　　金：癌基因及相关基因国家重点实验室开放基金资助项目(80-9)(1683000023)

摘　　要：目的:了解与细胞周期素A1互作的一个EST的组成结构和蛋白质细胞定位。方法:通过酵母双杂交试验获得了一个与细胞周期素A1互作的新EST,进一步获得了全长基因foca1,其读码框长度为666 bp,编码221个氨基酸。将该基因克隆进杆状病毒AcNPV及真核表达质粒pcDNA3中,分别进行了表达和定位。结果:获得了表达该基因的重组杆状病毒和融合了EGFP的真核表达质粒。蛋白印迹试验证明该基因在昆虫Tn5细胞中得到了表达;并发现该基因的产物在COS-7细胞中定向于细胞核内。结论:该蛋白与cyc lin A1(核蛋白)存在互作的可能性。此结果为深入研究foca1基因的功能打下了基础。Objective： To understand the structure and location of an EST protein interacting with cyclin A1. Methods： By yeast triple-hybrid test, a new EST（focal ） was acquired, which might be a candidate interacting with cyclin A1. The focal, a span gene, was obtained in further by RACE, with a length of ORF was 666 base pairs, encoding 221 amino acids. This gene was cloned into baculovirus AcNPV and pcDNA3 plasmid fusing to EGFP and then was expressed and located, respectively. Results： The recombinant baculovirus and plasmid were obtained. It demonstrated that the gene was expressed in insect Tn5 cells by Western-blot. The gene products was located in the nucleus of COS-7 cells tranfected with the recombinant plasmid pcDNA3-egfp-focal. Conclusion： It indicated that focal had the interacting probability with Cyclin A1 which also was a nuclear protein. Further studies will reveal the function of the focal gene.

关 键 词：FOCAL 细胞周期素 克隆 表达 杆状病毒 

分 类 号：Q789[生物学—分子生物学]