翅碱蓬CMO基因克隆、测序及植物表达载体的构建

中国生物工程杂志

研究简报

翅碱蓬CMO基因克隆、测序及植物表达载体的构建

仲崇斌刘长江费腾袁晓东孙丽慧

东北大学理学院生物技术研究所（沈阳）

Cloning and Sequencing of Suaeda hetroptera Kitag CMO cDNAand Construction of its recombinant Plant Expression Vector

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摘要： 从翅碱蓬叶片中提取总RNA，根据相关同源序列设计引物，使用反转录试剂盒进行RT-PCR，扩增得到翅碱蓬胆碱单加氧酶（CMO）cDNA，进行PCR产物测序；然后将CMOcDNAT-A克隆至pMD18-T-simple载体上，经测序正确后亚克隆至pBI121植物表达载体，进行酶切鉴定及克隆测序。

关键词： 胆碱单加氧酶 pMD18-T-simple载体; pBI121植物表达载体大肠杆菌JM109

Abstract: Total RNA was extracted from leaf of Suaeda hetroptera Kitag, then the CMO(choline monooxygenase)cDNA was amplified using the Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) method and cloned into pMD-T-simple vector. The positive clones from the Blue/White Screen were sequenced. After confirming its validity, we subcloned the CMO gene fragment into pBI121 vector. Double enzymes restriction and PCR analysis indicated that the pBI121/CMO recombinant plasmid was successfully constructed.

Key words: choline monooxygenase pMD18-T-simple Vector pBI121 Plant Expression Vection E.coli JM109

收稿日期: 2005-11-29 出版日期: 2006-07-25

通讯作者: 仲崇斌

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引用本文:

仲崇斌,刘长江,费腾,袁晓东,孙丽慧. 翅碱蓬CMO基因克隆、测序及植物表达载体的构建[J]. 中国生物工程杂志, .

. Cloning and Sequencing of Suaeda hetroptera Kitag CMO cDNAand Construction of its recombinant Plant Expression Vector. China Biotechnology, .

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2006/V26/I07/80

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