水稻条纹病毒CP与叶绿体Rubisco SSU引导肽融合基因的构建及原核表达

PCR扩增获得水稻叶绿体Rubisco SSU引导肽基因和水稻条纹病毒（Rice stripe virus，RSV）外壳蛋白（coat protein，CP）基因。分别借助于pGEX-4T-1和pET-29a表达载体，构建获得了融合基因PR-CP和PR-S-CP。测序鉴定融合基因构建正确后，将重组子pGEX-PR-CP和pET-PR-S-CP转化大肠杆菌BL21(DE3)，IPTG诱导表达后，利用SDS-PAGE电泳和Western blot对表达产物进行了鉴定。结果发现，融合基因表达产物大小与预期结果一致，且能与RSV CP抗体特异结合。说明融合基因中的RSV CP获得了正确表达，在融合蛋白中保持了自身独立的抗原活性，推断融合基因在生物体内的表达可能都能保持每个蛋白各自独立的结构和功能。

Construction of Fusion Gene RSV CP and Rubisco SSU Leader Peptide of Rice Chloroplast and Its Prokaryotic Expression

The coat protein (CP) gene of Rice stripe virus (RSV) and Rubisco SSU leader peptide gene of rice chloroplast were amplified by PCR。Depending on the expression vector pGEX-4T-1 and pET-29a, fusion gene PR-CP and PR-S-CP was constructed. The two fusion genes were confirmed correct by sequencing and analysis. The recombinant pGEX-PR-CP and pET-PR-S-CP were transformed into E. coli BL21 (DE3), and then induced to express by IPTG. The expression products of fusion gene were identified by SDS-PAGE electrophoresis and Western bolt. The result suggested that the fusion proteins were the same as we expected and could be specifically recognized by the antibody to RSV CP. This suggested that RSV CP gene in the fusion gene expressed correctly and maintained its antigen activity, so we could conclude that the expression products of fusion gene in the organism maybe could maintain its structure and function separately.