摘要
目的:copines蛋白家族是新近发现的一类分布广泛、进化保守的Ca2+依赖性磷脂结合蛋白,copineⅤ是其成员之一。为研究copineⅤ基因的功能,构建了人copineⅤ基因的真核表达质粒并转染HEK293细胞,以进一步研究其功能。方法:构建人copineⅤ编码区序列的真核表达质粒pCDNA4/copineⅤ,脂质体法转染HEK293细胞。通过RT-PCR、克隆形成实验和Hochesst33342染色等方法观察copineⅤ基因对HEK293细胞生长的影响。结果:外源表达的人copineⅤ可以诱导HEK293细胞死亡,无法筛选到稳定克隆。结论:人copineⅤ可以诱导HEK293细胞死亡,为进一步研究copineⅤ基因的生物功能提供了线索。
Objective： To study the function of copine V gene, a member of a novel family of ubiquitous Ca^2＋-dependent, phospholipid-binding proteins that have been conserved through evolution. Methods： The expression vector of copineV gene was constructed and its effect on HEK293 cell line investigated using RT-PCR, transient transfection, colne formaion assay, and Hochesst33342 dyeing assay. Results： Overexpression of copine V genes alters nuclear and cellular morphology of HEK2.93 cells and causes cell death. Conclusion： The overexpression of copine V gene in HEK2.93 cells can causes cell death.
出处
《生物技术通讯》
CAS
2006年第2期146-148,共3页
Letters in Biotechnology
基金
国家高技术研究发展计划课题(2002BA711A01-03)
