摘要
目的:研究人类精子膜表面是否存在电压依赖性阴离子通道(VDAC),并研究其生物学特性。方法:设计VDAC的特异性引物,从睾丸cDNA文库中用PCR技术扩增该通道的基因产物,用分子克隆技术体外表达重组VDAC的蛋白质。从精子表面用1%TritonX-100提取及氯仿/甲醇分离疏水性的膜表面蛋白,用免疫印迹法检测精子表面是否存在天然VDAC蛋白质。结果:人类睾丸cDNA文库含有VDAC的基因表达,人类精子膜表面发现靠N端α螺旋连接在精子膜上的VDAC蛋白质。结论:人类精子膜表面镶嵌VDAC蛋白质,该通道是以α螺旋连接在精子膜上,调控精子膜内外离子的转运和信号的转导,对精子的运动和功能至关重要。
Objective： To further study gene expression and characterization of voltage-dependent anion channels（VDACs） on human spermatozoa. Methods： VDACs were cloned by PCR from the testis cDNA library. Recombinant human sperm VDACs were produced in E. coil system by molecular cloning technology. Sperm membrane protein was extracted by 1% TritonX-100 and separated by chloroform/methanol. Results： The gene expression of VDACs was found in the human testis cDNA library and VDAC protein was detected located on the sperm membrane by a-helix. Conclusion： VDAC proteins, abundant on the human sperm membrane and responsible for anion transportation, play an important role in sperm signaling transduction and fertility.
出处
《中华男科学杂志》
CAS
CSCD
2007年第6期498-501,共4页
National Journal of Andrology
基金
国家"十一五"科技支撑计划(2006BAI03B12)
