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.
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