目的:为进一步研究人牛精浆蛋白(bovine sem inal p lasm a,BSP)相关蛋白(hum an BSP-related prote in,HBRP)的生物学功能,建立稳定表达的细胞系。方法:利用重组质粒pEGFP-C2/HBRP转染人胚肾HEK293细胞,通过G418筛选出药物抗性克隆,并进一步进行培养、传代。结果:建立了新基因HBRP稳定表达的基因工程细胞系。结论:实现了利用真核细胞系表达HBRP蛋白,为该蛋白功能的研究奠定了基础,对该基因功能的进一步研究有重要的意义。
Objective To investigate the biology function of novel protein related to bovineseminal plasma protein in human testis.Methods Recombination pcDNA3/HBRP was constructed and transfected to HEK293cell and permanently expression cell line was established. The activity of proteinkinase C (PKC) of the cell line was detected by autoradiography method.Results The stable expression cell line of HBRP was obtained. The HBRP inhibitedthe activity of PKC significantly.Conclusion One of the new functions of novel sperm binding protein in human is theinhibitor action on activity of PKC. It may be involved in the sperm capacitation andacrosome reaction.
Jie ZHANG Ying LIU Wen-hui SU Guo-li WANG Zhi-hong ZONG Bing-zhi YU
目的:探讨小鼠卵母细胞不同细胞周期时相中,由于p34cdc2不同磷酸化状态所引起的电泳迁移率变化。方法:用免疫印迹(W estern b lot)技术观察G2期和MⅡ期小鼠卵母细胞中p34cdc2电泳迁移率的变化。结果:p34cdc2在G2期只表现两条带即上、中带;而MⅡ期表现为中、下带。结论:可以用电泳迁移率的变化作为观察p34cdc2磷酸化状态的一种方法。
