| Kringle 5的真核植物细胞穿梭表达载体的构建 |
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| 引用本文： | 郑合明,薛乐勋.Kringle 5的真核植物细胞穿梭表达载体的构建[J].郑州大学学报(医学版),2002,37(2):156-159. |
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| 作者姓名： | 郑合明 薛乐勋 |
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| 作者单位： | 1. 郑州大学医学实验中心细胞生物学研究室,郑州,450052;河南省卫生防疫站,郑州,450003
2. 郑州大学医学实验中心细胞生物学研究室,郑州,450052 |
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| 基金项目： | 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 |
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| 摘 要： | 目的：构建含有目的基因kring5的真核植物细胞穿酸表达载体。方法：应用RT－PCR方法，从正常人肝脏组织中获得kringle5基因，测序后，再通过DNA重组技术，将kringle5基因片段克隆到真核植物细胞表达载体pBI121和pCAMBIA3301上。结果：得到的kringle5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，而且可以在植物细胞中表达。
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| 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 |
| 修稿时间： | 2001年8月8日 |
Construction of shuttle expression vector in eukaryocyte plant cells |
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| ZHENG Heming ,XUE Lexun Cell Biology Laboratory,Center of Medical Experiments,Zhengzhou University,Zhengzhou Anti Epidemic and Health Station of Henan Province,Zhengzhou.Construction of shuttle expression vector in eukaryocyte plant cells[J].Journal of Zhengzhou University: Med Sci,2002,37(2):156-159. |
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| Authors: | ZHENG Heming XUE Lexun Cell Biology Laboratory Center of Medical Experiments Zhengzhou University Zhengzhou Anti Epidemic and Health Station of Henan Province Zhengzhou |
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| Affiliation: | ZHENG Heming 1,2),XUE Lexun 1) 1)Cell Biology Laboratory,Center of Medical Experiments,Zhengzhou University,Zhengzhou 450052 2)Anti Epidemic and Health Station of Henan Province,Zhengzhou 450003 |
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| Abstract: | 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. |
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| Keywords: | angiostatin kringle 5 shuttle expression vector RT PCR promoter CaMV 35S |
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