中文摘要：

目的：克隆小鼠Bmi1 cDNA并进行原核表达，为下一步制备BMI1多克隆抗体奠定基础。方法：从小鼠睾丸组织中克隆Bmi1 cDNA，测序并利用BLAST程序比对证明该基因序列正确后，构建原核表达载体pET-28c-Bmi1，将其转人大肠埃希菌BL21中，异丙基-B-D-硫代半乳糖苷（IPTG）诱导表达并通过免疫印迹检测重组目的蛋白。结果：成功克隆了小鼠Bmi1 cDNA，原核表达的融合蛋白可与His标签抗体和BMI1单克隆抗体特异性结合。结论：小鼠睾丸组织中Bmi1基因有表达；成功克隆了小鼠Bmi1 cDNA并在大肠埃希菌BL21中正确表达了重组蛋白rBMI1。

英文摘要：

Objective： To produce BMI1 polyclonal antibody, mouse Bmi1 cDNA was cloned from mouse testis and expressed in E. coli BL21. Methods： Broil gene was amplified from mouse testis by RT-PCR and inserted into the prokaryotic expression vector pET- 28c（ ＋ ）. Subsequently the recombined vector was transformed and expressed in E. coli BL21 （DE3） and the immunogenicity of recombined protein BMI1 （ rBMI1 ） was tested by Western blot. Results ： Mouse Broil cDNA of 975 bp was successfully cloned and recombined. E. coli BL21 strains expressed rBMI1 were screened. The expression protein amounted to 12% of the total bacterial protein after induced with IPTG, which included inclusion body and soluble protein. Inclusion body was the major pattern of the expression that amounted to 71% of the insoluble protein. Western blot analysis showed that rBMI1 could be specially recognized by mouse monoclonal IgG1 anti-BMI1 and His-tag antibody. Conclusion ： There was expression of Broil gene in mouse testis. Mouse Bmi1 cDNA was successfully cloned and expressed prokaryoticly.