| 番木瓜环斑病毒南瓜分离物外壳蛋白基因的克隆及序列分析 |
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| 引用本文： | 杨国慧,张仲凯,崔崇士.番木瓜环斑病毒南瓜分离物外壳蛋白基因的克隆及序列分析[J].植物保护学报,2007,34(3):273-276. |
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| 作者姓名： | 杨国慧 张仲凯 崔崇士 |
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| 作者单位： | 云南省农业生物技术重点实验室，云南省农业生物技术重点实验室，东北农业大学园艺学院 昆明650223东北农业大学园艺学院，哈尔滨150030，昆明650223，哈尔滨150030 |
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| 摘 要： | 利用电镜和酶联免疫法在云南省采集到的5份南瓜病样中检测到番木瓜环斑病毒(Papayaring spot virus,PRSV)。为了进一步从分子水平确定云南省南瓜病毒病原种类,并为下一步转基因育种提供抗性基因,采用反转录PCR(RT-PCR)方法扩增了5个分离物的外壳蛋白(coat protein,CP)基因片段,并克隆到pGEM-T载体中。核苷酸序列测定表明,番木瓜环斑病毒石屏分离物(PRSV-SP)和番木瓜环斑病毒蒙自分离物(PRSV-MZ)的CP基因长873nt,编码290个氨基酸,番木瓜环斑病毒峨山分离物(PRSV-ES)、番木瓜环斑病毒版纳分离物(PRSV-BN)和番木瓜环斑病毒宾川分离物(PRSV-BC),3个分离物CP基因长867nt,编码288个氨基酸。PRSV5个分离物核苷酸序列的同源性在94%以上,氨基酸序列的同源性在96%以上。与国内外17个分离物相比,核苷酸序列同源性为89.6%~98.7%,氨基酸序列同源性为86.5%~99.6%。其中PRSV-SP和来自于越南分离物PRSV-V47无论是核苷酸序列,还是氨基酸序列同源性都达到了最高,而5个分离物与来自于巴西(PRSV-BR)、美国(PRSV-USA)、墨西哥(PRSV-Y)核苷酸序列同源性均低于90%。
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| 关 键 词： | 南瓜 番木瓜环斑病毒 外壳蛋白基因 序列分析 |
| 收稿时间： | 2006/10/12 0:00:00 |
| 修稿时间： | 2006-10-12 |
Cloning and sequence analysis of CP genes of PRSV from pumpkin |
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| YANG Guo-hui,ZHANG Zhong-kai and CUI Chong-shi.Cloning and sequence analysis of CP genes of PRSV from pumpkin[J].Acta Phytophylacica Sinica,2007,34(3):273-276. |
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| Authors: | YANG Guo-hui ZHANG Zhong-kai and CUI Chong-shi |
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| Institution: | 1. Yunnan Key Lab of Agro-Biotechnology, Kunming 650223, Yunnan Province, China; 2. Horticultural College of Northeast Agricultural University, Harbin 150030, Heilongjiang Province, China |
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| Abstract: | Papaya ring spot virus(PRSV)was tested with electron microscopy and ELISA from 5 diseased pumpkin samples collected in Yunnan Province.To further determine pathogen types of pumkin viral disease in Yunnan Province on a molecular level and provide anti-disease genes for trans-gene breeding,CP gene fragment was amplified by RT-PCR from 5 isolates and cloned to pGEM-T vector.Nucleotide sequence showed that CP gene of PRSV Shiping isolate(PRSV-SP)and PRSV Mengzi isolate(PRSV-MZ)contains 873 nt encoding 290 amino acid,while that of PRSV Eshan isolate(PRSV-ES),PRSV Banna isolate(PRSV-BN)and PRSV Binchuan isolate(PRSV-BC)contains 867 nucleotide encoding 288 amino acid.The nucleotide and amino acid homology of the 5 isolates is higher than 94% and 96% respectively.Compared with sequence of other 17 foreign and domestic isolates,the homology of nucleotide is between 89.6% and 98.7%,the homology of amino acid is between 86.5% and 99.6%.It was also revealed that PRSV-SP was shared highest similarity with sequence of isolate PRSV-V47 from Vietnam,but these five isolates were shared low similarity with sequence of isolate PRSV-BR from Brazil,PRSV-USA from USA and PRSV-Y from Mexico,among which the homology of nucleotide is less than 90%. |
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| Keywords: | Pumpkin Papaya ring spot virus CP gene sequence analysis |
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