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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。
Recombinant plant expression vector containing HPV45-L1 and HPV58-L1 gene was constructed by using rice endosperm cell bioreactor, which provided a new efficient and low-cost expression pattern for HPV-45L1 and HPV58-L1 protein. The coding genes of human papillomavirus HPV45-L1 and HPV58-L1 were cloned by PCR and recombined into the intermediate vector pMP3 and the plant expression vector pCAMBIA-1300 to construct the plant expression vector pCAMBIA containing the HPV45-L1 and HPV58-L1 genes -1300-pMP3-HPV45-L1, pCAMBIA-1300-pMP3-HPV58-L1, followed by transformation of rice plants mediated by Agrobacterium tumefaciens-mediated rice callus. HPV-L1 transgenic rice plants were obtained. PCR results showed that HPV45-L1 and HPV58-L1 genes have been integrated into rice genome, indicating that rice endosperm-specific expression vector has been successfully constructed.