[黄瓜花叶病毒cDNA载体在烟草细胞中表达外源基因]

[Cucumber mosaic virus cDNA vector expressed foreign genes in tobacco cells].

作者信息

Lei W L, Fang R X, Zhang G H

机构信息

Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2001 Jan;17(1):40-5.

PMID:11330185

Abstract

Full-length cDNA of SD strain CMV (SD-CMV) RNA 3 was cloned and sequenced. An Nsi I site was created at the sequence around the start codon of coat protein (CP) gene and a replacement cassette was constructed. The CP gene was replaced by green fluorescent protein (GFP) gene, beta-glucuronidase (GUS) gene or mouse dihydrofolate reductase (DHFR) gene, respectively. The cDNAs of Fny-CMV RNAs 1 and 2 and the chimeric SD-CMV RNA 3 were cloned between the 35S promoter and terminator separately. Tobacco protoplasts transfected with the CMV cDNA vectors expressed the three reporters, implying that CMV could be used as an expression vector.

摘要

克隆并测序了SD株黄瓜花叶病毒（SD-CMV）RNA 3的全长cDNA。在外壳蛋白（CP）基因起始密码子周围的序列处创建了一个Nsi I位点，并构建了一个替换盒。CP基因分别被绿色荧光蛋白（GFP）基因、β-葡萄糖醛酸酶（GUS）基因或小鼠二氢叶酸还原酶（DHFR）基因取代。Fny-CMV RNA 1和2的cDNA以及嵌合的SD-CMV RNA 3分别克隆在35S启动子和终止子之间。用CMV cDNA载体转染的烟草原生质体表达了这三种报告基因，这意味着CMV可作为一种表达载体。