摘要
目的研究小鼠Boule蛋白与精子成熟相关基因Crem之间的相互作用,揭示Boule蛋白在精子发生过程中所起的作用。方法构建pGEX-Boule表达载体,将表达载体转化宿主菌(BL21)感受态细胞,诱导纯化出Boule蛋白,应用RT-PCR和凝胶滞后实验(EMSA)分析与Boule蛋白结合的mRNA。结果表达载体pGEX-Boule构建成功,纯化出GST-Boule融合蛋白,RT-PCR和EMSA实验证明Boule蛋白能够结合Crem3′UTR。结论 Boule蛋白能够结合Crem3′UTR,可能通过调控Crem蛋白的表达作用于精子发生过程。
Objective To investigate the interaction between Boule protein and sperm maturation related gene Crem and reveal the function of Boule protein in spermatogenesis.Methods The pGEX-Boule expression vector was constructed and transformed into BL21 competent cells,and induced Boule protein was purified.RT-PCR and EMSA were performed to analyze the mRNA bond to the Boule protein.Results The recombinant expression vector pGEX-Boule was successfully constructed and purified GST-Boule protein was obtained,RT-PCR and EMSA experiments indicated that Boule protein was capable to bind to Crem3′UTR.Conclusion Boule protein may regulate the expression of the Crem protein by binding to Crem3′UTR and plays a role in spermatogenesis process.
出处
《重庆医学》
CAS
CSCD
北大核心
2012年第24期2462-2464,2468,共4页
Chongqing medicine
基金
国家重点基础研究发展计划(973计划)基金资助项目(2012CB947600)
国家自然科学基金资助项目(31070676)
教育部春晖计划基金资助项目(Z2009-1-61003)
