抑癌基因OVCA1 A34D突变对其生物半衰期的影响  

Mutation of tumor-suppressor OVCA1 A34D extends the half-life of the protein

作　　者：魏巍[1] 李岩[1] 石敏[1] 张莹[1] 吕倩[1] 郭娜[1] 赵春艳[1] 

机构地区：[1]大连医科大学检验医学院,辽宁大连116044

出　　处：《中国微生态学杂志》2016年第7期778-780,共3页Chinese Journal of Microecology

基　　金：辽宁省自然科学基金(2013023013)

摘　　要：目的为探索抑癌基因OVCA1结构与功能的关系,阐明其在肿瘤发生发展中的作用机制,本研究构建OVCA1A34D突变体并探讨该位点突变对其生物半衰期的影响。方法以本实验室制备并保存的含有人OVCA1基因全长序列的质粒为模板,采用分子克隆方法构建了GFP-tagged-OVCA1A34D突变体重组质粒,并经测序证实。应用脂质体法将突变体质粒转染入体外培养细胞,观察OVCA1A34D突变体蛋白在细胞中的表达。采用蛋白合成抑制剂放线菌酮抑制蛋白合成后检测OVCA1A34D基因突变对其蛋白生物半衰期的影响。结果成功构建了GFP-tagged-OVCA1A34D突变体重组质粒,并在细胞中成功表达。OVCA1A34D突变体蛋白的半衰期与野生体相比明显延长。结论 OVCA1A34D位点突变可导致其生物半衰期延长。Objective To explore the relationship between the structure and function of tumor suppressor OVCA1, in order to clarify the mechanism of OVCA1 in tumorigenesis and progression. Methods The GFP-tagged-OVCA1 A34D mutant was built by gene cloning and was confirmed by sequencing. The mutant was transfected into cells with liposome assay. The expression of OVCA1 A34D mutant protein was detected and the half-life of the protein was measured by inhibiting protein synthesis with cycloheximide （CHX）. Results GFP-tagged-OVCA1 A34D was established successfully and was expressed in cells. The half-life of OVCA1 A34D mutant protein was longer compared with that of wild type OVCA1 protein. Conclusion OVCA1 A34D mutation extends the protein′s half-life.

关 键 词：OVCA1 点突变 半衰期 

分 类 号：R733[医药卫生—肿瘤]