东方山羊豆GoMIPS双元表达载体的构建及鉴定

李春艳,刘建宁,高洪文

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家畜生态学报 ›› 2013, Vol. 34 ›› Issue (1) : 25-28.
科学研究

东方山羊豆GoMIPS双元表达载体的构建及鉴定

  • 李春艳,刘建宁,高洪文
作者信息 +

Construction and Indentification of Binary Expression Vector ofGalega Orientalis GoMIPS GeneLI

  • LI Chun-yan,LIU Jian-ning,GAO Hong-wen
Author information +
History +

摘要

从东方山羊豆(Galega.orientalisL.)中克隆出的GoMIPS(肌醇-1-磷酸合成酶)基因序列设计合成1对带有NcoI和SpeI酶切位点的引物,用RT-PCR方法从东方山羊豆幼嫩叶片总RNA中扩增出GoMIPS基因的cDNA序列,经纯化后,克隆至pMD19-T载体上,构建pMD-MIPS的中间载体,测序鉴定。然后用NcoI和SpeI限制性酶对含有目的基因的pMD-MIPS和pCAMBIA1302空载体进行双酶切,通过酶切鉴定和测序分析表明,已成功构建了CaMV35S启动子驱动GFP报告基因的双元植物表达载体pCAMBIA1302-MIPS。

Abstract

A pair of primers containing the restriction sites for NcoI/SpeI were designed according to the cloned GoMIPS (inositol-1 - phosphate synthase) gene sequence of Galega orientalis. The GoMIPS cDNA was amplified by RT-PCR from the total RNA of the young leaves of the Oriental Galega. After purification, RT-PCR products were cloned to the pMD19-T vector were dealt to construct the pMD-MIPS in the middle of the carrier DNA sequencing. pMD-MIPS containing the target gene and pCAMBIA1302 empty vector with NcoI / SpeI restriction enzyme digestion,the restriction endonuclease and sequence analysis shows that a dual CaMV35S promoter-driven GFP reporter geneplant expression vector pCAMBIA1302-MIPS has been successfully constructed.

关键词

东方山羊豆(Galega.orientalisL.) / GoMIPS / pMD19-T / pCAMBIA1302

Key words

Galega. orientalis L. / GoMIPS (inositol -1- phosphate synthase) / pMD19-T vector / pCAMBIA1302 vector

引用本文

导出引用
李春艳,刘建宁,高洪文. 东方山羊豆GoMIPS双元表达载体的构建及鉴定. 家畜生态学报. 2013, 34(1): 25-28
LI Chun-yan,LIU Jian-ning,GAO Hong-wen. Construction and Indentification of Binary Expression Vector ofGalega Orientalis GoMIPS GeneLI. Journal Of Domestic Animal Ecology. 2013, 34(1): 25-28

参考文献

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基金

山西省农业科学院畜牧兽医研究所基金项(YGG0853)

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