甘蔗花叶病毒福建分离物外壳蛋白基因的克隆及序列分析

姚伟, 段真珍, 周会, 何正权, 张木清, 陈如凯

植物病理学报 ›› 2006, Vol. 36 ›› Issue (4) : 378-381.

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植物病理学报 ›› 2006, Vol. 36 ›› Issue (4) : 378-381.
研究简报

甘蔗花叶病毒福建分离物外壳蛋白基因的克隆及序列分析

  • 姚伟1,2, 段真珍1, 周会2, 何正权1, 张木清2, 陈如凯2
作者信息 +

Cloning and sequence analysis of coat protein genes of Sugarcane mosaic virus isolates obtained from Fujian

  • YAO Wei1,2, DUAN Zhen-zhen1, ZHOU Hui2, HE Zheng-quan1, ZHANG Mu-qing2, CHEN Ru-kai2
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文章历史 +

摘要

A fujian isolate of Sugarcane mosaic virus named SCMV-FJ was isolated from infected sugarcane. Cloning and sequence analysis of the coat protein gene of this isolate was carried out. A pair of primers was designed and synthesized based on the nucleotide sequences of coat protein genes of sugarcane mosaic viruses reported. The coat protein gene of SCMV-FJ was amplified from the extracted total RNA of the infected sugarcane by using RT-PCR, and cloned into the pMD18-T vector. The sequencing result indicated that the cloned segment included a 1137 bp open reading frame(ORF) and a 228 bp 3' untranslated region, in which the ORF comprised the whole coat protein and part of the nuclear inclusion b. The nucleotide and the deduced amino acid sequences of the coat protein gene were compared with those of the other isolates or strains of SCMV subgroup reported in GenBank. The result showed that it shares 56.8%-97.1% and 55.3%-99.4% homology in nucleotide and the putative amino acid sequences, respectively, with the highest amino acid homology of 99.4% with SCMV-D. Thus it was identified as a SCMV-D. This experiment provided a rapid, sensitive and relatively inexpensive method for RT-PCR detection of SCMV. At the same time, the cloning of SCMV-FJ coat protein gene provided the foundation for plant gene engineering against SCMV.

Abstract

A fujian isolate of Sugarcane mosaic virus named SCMV-FJ was isolated from infected sugarcane. Cloning and sequence analysis of the coat protein gene of this isolate was carried out. A pair of primers was designed and synthesized based on the nucleotide sequences of coat protein genes of sugarcane mosaic viruses reported. The coat protein gene of SCMV-FJ was amplified from the extracted total RNA of the infected sugarcane by using RT-PCR, and cloned into the pMD18-T vector. The sequencing result indicated that the cloned segment included a 1137 bp open reading frame(ORF) and a 228 bp 3' untranslated region, in which the ORF comprised the whole coat protein and part of the nuclear inclusion b. The nucleotide and the deduced amino acid sequences of the coat protein gene were compared with those of the other isolates or strains of SCMV subgroup reported in GenBank. The result showed that it shares 56.8%-97.1% and 55.3%-99.4% homology in nucleotide and the putative amino acid sequences, respectively, with the highest amino acid homology of 99.4% with SCMV-D. Thus it was identified as a SCMV-D. This experiment provided a rapid, sensitive and relatively inexpensive method for RT-PCR detection of SCMV. At the same time, the cloning of SCMV-FJ coat protein gene provided the foundation for plant gene engineering against SCMV.

关键词

Sugarcane mosaic virus / coat protein / molecular detection

Key words

Sugarcane mosaic virus / coat protein / molecular detection

引用本文

导出引用
姚伟, 段真珍, 周会, 何正权, 张木清, 陈如凯. 甘蔗花叶病毒福建分离物外壳蛋白基因的克隆及序列分析[J]. 植物病理学报, 2006, 36(4): 378-381
YAO Wei, DUAN Zhen-zhen, ZHOU Hui, HE Zheng-quan, ZHANG Mu-qing, CHEN Ru-kai. Cloning and sequence analysis of coat protein genes of Sugarcane mosaic virus isolates obtained from Fujian[J]. Acta Phytopathologica Sinica, 2006, 36(4): 378-381

基金

国家“八六三”高技术研究发展计划资助项目(2002AA241031);湖北省教育厅青年基金资助项目(Q200513002)
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