Kringle5的真核植物细胞穿梭表达载体的构建  

Construction of shuttle expression vector in eukaryocyte plant cells

作　　者：郑合明[1] 薛乐勋[1] 

机构地区：[1]郑州大学医学实验中心细胞生物学研究室

出　　处：《郑州大学学报（医学版）》2002年第2期156-159,共4页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家"九五"科技攻关基金资助项目 96 90 1 0 5 189; 河南省重大科技攻关基金资助项目 0 12 2 0 3 2 5 0 0

摘　　要：目的 :构建含有目的基因kringle5的真核植物细胞穿梭表达载体。方法 :应用RT PCR方法 ,从正常人肝脏组织中获得kringle 5基因 ,测序后 ,再通过DNA重组技术 ,将kringle 5基因片段克隆到真核植物细胞表达载体pBI12 1和pCAMBIA330 1上。结果 :得到的kringle 5基因序列测定结果与文献相符 ,重组载体pB1k5和pC33k5经酶切鉴定正确 ,含有植物高效表达CaMV35S启动子。结论 :重组载体pB1k5和pC33k5不仅可以在大肠杆菌中稳定复制 。Aim:To construct the plant shuttle expression plasmids containing the target gene kringle 5. Methods: First, the cDNA was obtained from healthy human liver by use of RT PCR. Second, PCR technique was used to obtain target gene kringle 5, and its sequence was identified by DNA sequencing kit. Third, the shuttle expression plasmids were constructed by use of sub cloning technique. The target gene kringle 5 was cloned into plasmids pBI121 and pCAMBIA3301, respectively. Results: The recombinant plant expression plasmids pB1k 5 and pC33k 5 were identified to be correct, which contained plant expression promoter CaMV 35S. Conclusion: The recombinant plasmids pB1k 5 and pC33k 5 not only steadily replicate in E.coli cells, but also express in plant cells.

关 键 词：血管抑素 kringle5基因 穿梭表达载体 反转录PCR 启动子CaMV35S 真核植物细胞 

分 类 号：R459.9[医药卫生—治疗学] Q78[医药卫生—临床医学]