XUE Chao-Yang, ZHOU Xue-Ping, LIU Yong and LI De-Bao. Cloning,Sequence Analysis and Expression in E .coli of Coat Protein Gene and 3 Noncoding Region of Tom ato M osaic Virus[J]. Virologica Sinica, 1998, 13(2).
Citation: XUE Chao-Yang, ZHOU Xue-Ping, LIU Yong, LI De-Bao. Cloning,Sequence Analysis and Expression in E .coli of Coat Protein Gene and 3 Noncoding Region of Tom ato M osaic Virus .VIROLOGICA SINICA, 1998, 13(2) : 150.

番茄花叶病毒外壳蛋白基因及3′端非编码区的克隆、序列分析及其表达

  • 根据已报道的番茄花叶病毒L株系(ToMVL)序列人工合成引物,经RTPCR扩增并克隆了我国番茄花叶病毒分离物(ToMVS1)的外壳蛋白CP基因及3′端非编码区。序列测定结果表明,所得cDNA共长682个核苷酸,其中CP基因含480个核苷酸,编码158个氨基酸,3′端非编码区含202个核苷酸,其核苷酸序列与ToMVL株系具有99.5%的同源率。将该基因片段克隆到pGEMEX1载体中,转入E.coli后诱导表达,经Westernblot检测证明,该基因已在大肠杆菌中正确表达。这是我国首次报道ToMVCP基因序列。

Cloning,Sequence Analysis and Expression in E .coli of Coat Protein Gene and 3 Noncoding Region of Tom ato M osaic Virus

  • The coat protein (CP) gene and 3′noncoding region of tomato mosaic virus Sl isolate (ToMV Sl) were amplified by RT PCR with specific primers designed on the basis of reported ToMV L strain (ToMV L) sequence. The cDNA fragment was cloned and sequenced,its sequence showed that the CP gene has 480 nucleotides in length, which encodes 158 amino acids and 3′ noncoding region contains 202 nucleotides. The nucleotide sequence of ToMV Sl showed 99.5% homology when compared with that of ToMV L and 76.2% homology when compared with that of TMV Ul. The CP gene was inserted into pGEMEX 1, Western immuno blot showed that this gene was expressed in E.coli with the proper reading frame. The gene was also inserted into plant expression vector pBI121.

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    Cloning,Sequence Analysis and Expression in E .coli of Coat Protein Gene and 3 Noncoding Region of Tom ato M osaic Virus

    • 1. 1nstitute of Biotechnology.Zhejiang Agricultural University,Hangzhou 310029

    Abstract: The coat protein (CP) gene and 3′noncoding region of tomato mosaic virus Sl isolate (ToMV Sl) were amplified by RT PCR with specific primers designed on the basis of reported ToMV L strain (ToMV L) sequence. The cDNA fragment was cloned and sequenced,its sequence showed that the CP gene has 480 nucleotides in length, which encodes 158 amino acids and 3′ noncoding region contains 202 nucleotides. The nucleotide sequence of ToMV Sl showed 99.5% homology when compared with that of ToMV L and 76.2% homology when compared with that of TMV Ul. The CP gene was inserted into pGEMEX 1, Western immuno blot showed that this gene was expressed in E.coli with the proper reading frame. The gene was also inserted into plant expression vector pBI121.

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