HPV45-L1和HPV58-L1蛋白植物表达载体的构建及水稻转基因植株的鉴定

Construction of HPV45-L1 and HPV58-L1 Plant Expression Vectors and Identification of Transgenic Rice Plants

利用水稻胚乳细胞生物反应器,构建含有HPV45-L1和HPV58-L1基因的重组植物表达载体,为HPV-45L1和HPV58-L1蛋白的表达提供一种新的高效、低廉的表达方式。利用PCR技术克隆人乳头瘤病毒HPV45-L1和HPV58-L1蛋白编码基因,将其重组于中间载体pMP3和植物表达载体pCAMBIA-1300中,构建含HPV45-L1和HPV58-L1基因的植物表达载体pCAMBIA-1300-pMP3-HPV45-L1、pCAMBIA-1300-pMP3-HPV58-L1,随后采用根癌农杆菌侵染水稻愈伤组织介导的水稻遗传转化,获得HPV-L1转基因水稻植株。PCR检测结果表明,HPV45-L1和HPV58-L1基因已整合到水稻基因组中,说明已成功构建水稻胚乳特异性表达载体。

The rice seeds were used as the cell bioreactor to construct the recombinant plant expression vector containing HPV45-L1 and HPV58-L1 genes to provide a new efficient and inexpensive method for HPV45-L1 and HPV58-L1 protein expression.The HPV45-L1 and HPV58-L1 coding sequence（ORF）was amplified by PCR and then subcloned into middle vector pMP3 and plant expression vector pCAMBIA-1300 to construct the rice endosperm-specific expression vector pCAMBIA-1300-pMP3-HPV45-L1 and pCAMBIA-1300-pMP3-HPV58-L1.Subsequently,the transgenic rice plants containing HPV-L1 ORF gene were obtained by agrobacterium-mediated genetic transformation of rice callus.The PCR test showed that the HPV45-L1 and HPV58-L1 genes had integrated into the rice genome,suggesting the successful construction of rice endosperm specific expression vectors.