Nucleotide sequence analysis and detection application of the coat protein gene of Sweet potato latent virus isolated from central Taiwan

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题名:	Nucleotide sequence analysis and detection application of the coat protein gene of Sweet potato latent virus isolated from central Taiwan
作者:	Wang, L.-Y.;Chen, K.-C.;Chen, T.-C.;Yeh, S.-D.
贡献者:	Department of Biotechnology
关键词:	?藷潛伏病毒,核酸選殖,序列分析,檢測;Sweet potato latent virus,gene cloning,sequence analysis,detection
日期:	2007
上传时间:	2010-03-15 08:12:21 (UTC+0)
出版者:	Asia University
摘要:	於嘉義地區，自呈現黃斑及葉脈斑駁的甘藷葉片可分離到似potyvirus的長絲狀病毒(potyvirus-like isolate CY)。利用對應Potyvirus屬病毒基因體核糖核酸3'端之簡併式引子及寡合腺嘌呤引子，可由呈現局部斑之奎藜總量核糖核酸(total RNA)以反轉錄聚?連鎖反應增幅出約1.9-kb核酸片段。此核酸片段經選殖及核酸序列分析，全長含1880個核?酸序列。由5'端起此序列對應的胺基酸共561個，分別?268個胺基酸於細胞核內含體b(nuclear inclusion b, NIb)、及293個胺基酸之全長度鞘蛋白(coat protein CP)基因。3'端非轉譯區(3' non-coding region, 3-NCR)則由197個核酸組成。兩者之間蛋白?切位落於VHHQ/A之間，且由鞘蛋白N端起第79個胺基酸位置具有被蚜蟲傳播能力之DAG序列，不同於SPLV-TW之DTG。與登錄於GenBank之SPLV分離株比對核酸序列，與親緣最近之SPLV-TW病毒其CP及3'-NCR區域之核?酸序列相同度分別有96.5%及100%。鞘蛋白基因演化關係分析此病毒和台灣及中國大陸分離株較?親近而與日本分離秣較?疏遠。根據解得SPLV-CY鞘蛋白基因序列，設計之專一性引子對L166/L841，可由SPLV-CY感染之奎藜、煙草及目藷等病株檢測出專一性675 bp核酸片段。 A potyvirus-like agent (isolate CY) was isolated from sweet potato with leaf symptoms of chlorotic spots and vein mottling in Chia-Yi area, Taiwan. A 1.9-kb DNA product was amplified by reverse transcription polymerase chain reaction (RT-PCR) from infected tissues of C. quinoa using the oligo (dT) and degenerate primers for potyviruses. cloned, and then sequenced. The cDNA fragment reflected 1880 nucleotides (nts) corresponding to the 3'-terminal region of a potyvirus and the sequence comparison indicated that the isolate CY belongs to Stint potato latent virus (SPLV). The deduced amino acid sequence was determined as 561 residues that contains a part of the Y terminal region of NIb gene (268 residues) and the complete .sequence of coat protein (CP) gene (293 residue). The 3-terminal region of the cDNA was determined containing a non-coding region (NCR) of 197 nts. The DAG triplet for aphid transmissibility was found at the 7-9 residues from the N terminus of CP gene. Multiple sequence alignment of the known sequences of SPLV indicated that the CP gene and the NCR of CY isolate share 96.5 and l00% nucleotide identities, respectively, with those of SPLV-TW. The phylogenetic analysis indicated that the CY isolate is closely related to Taiwan and China isolates, but distantly related to a Japan isolate. When a pair of specific primers designed from the CP gene of SPLV-CY was used for detection by RT-PCR, a 675 bp specific DNA fragment was amplified rout SPLV-CY infected plants.
關聯:	Plant Pathology Bulletin 16:141-148
显示于类别:	[生物科技學系] 期刊論文

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