小鼠B7-2基因的克隆、测序及其重组质粒的构建

目的：克隆小鼠B7-2基因编码区的cDNA序列，并构建其表达载体。方法：利用逆转录聚合酶链反应（RT-PCR）法，以小鼠脾细胞mRNA为模板，扩增获得B7-2基因，与pMD-18T连接做全自动测序，并利用基因重组技术构建包含B7-2基因的表达质粒pcDNA3.1-CMV-B7-2及pcDNA3.1-Egr-B7-2。结果：经测序证实获得的B7-2基因与文献报道的基本一致，并成功地构建了表达质粒。结论：成功克隆了B7-2的编码基因，并成功构建了表达载体pcDNA3.1-CMV-B7-2及pcDNA3.1-Egr-B7-2。

Cloning and sequencing of mouse B7-2 gene and construction of its recombinant plasmids

Objective To clone the sequence of the cDNA of mouse B7 2 gene and construct its expression vectors. Methods Using mouse splenocyte mRNA as template to obtain full length B7 2 with the technique of RT PCR followed by automatic sequencing of pMD18T B7 2 and to construct recombinant plasmids containing CMV, Egr 1 and B7 2 gene with recombinant DNA technique. Results Sequencing proved the cloned B7 2 cDNA to be essentially identical with that reported in the literature and the recombinant plasmids containing CMV, Egr 1 and B7 2 gene were constructed successfully. Conclusion B7 2 cDNA was successfully cloned and two expression vectors pcDNA3 1 CMV B7 2 and pcDNA3 1 Egr B7 2 were constructed.