Sequence Analysis of Two Membrane-Associated Protein Genes of a Porcine Reproductive and Respiratory Syndrome Virus, Taiwan MD-001 Strain

豬生殖與呼吸綜合症病毒台灣MD-001分離株基因之選殖與序列分析：E與M兩個膜相關蛋白

闕玲玲(Ling-Ling Chueh) ； 李泔弘(Kan-Hung Lee)

《中華民國獸醫學會雜誌》 27卷2期 (2001/06) Pp. 80-88

https://doi.org/10.7009/JTCSVS.200106.0080

摘要

本實驗以1993年分離自本省豬生殖與呼吸綜合症病毒台灣MD-001分離株進行病毒結構基因之選殖。以特異性引子自病毒基因體經反轉錄與PCR反應得到病毒之cDNA，將所得片段經核酸定序發現此段DNA包含有兩個開放讀碼區各含603與525核苷酸。進一步將此序列與國外分離株進行比對證實此二基因分屬豬生殖與呼吸綜合症病毒ORF5（E基因）與ORF6（M基因），而此E與M兩基因間有16個核苷酸重疊。將台灣MD-001分離株之E基因核苷酸序列與歐洲分離株(Lelystad, Boxmeer, PRRS-OLOT)相較發現其相似性為52.1-53.7%；M基因之相似性則為63.8-64.8%。當與美國株(VR2332, VR2385, 16244B, MN1)，加拿大株(IAF-Exp91)，日本株(EDRD-1)與中國株(CH-1a)株進行比對時則發現相似性驟然提高許多：E基因83.1-88.7%, M基因91.6-94.5%。將兩核苷酸序列翻譯為氨基酸後發現E與M兩蛋白皆富含疏水性區域（分別佔E蛋白之42.5%與M蛋白之43.7%），此特性與兩蛋白皆為膜蛋白一致。

關鍵字

豬生殖與呼吸綜合症病毒 ； 台灣MD-001分離株 ； E與M蛋白 ； 序列分析

並列摘要

Two membrane associated protein genes, a putative glycosylated envelope protein (E) and an unglycosylated membrane protein (M), of porcine reproductive and respiratory syndrome virus (PRRSV) from a local isolate (MD-001 strain) were cloned and analyzed. After priming with specific oligonucleotides, the cDNA covering the E and M genes of the PRRSV were copied. The sequencing results of the obtained cDNA clones revealed two open reading frames (ORFs) which consisted of 603 and 525 nucleotides, respectively. A comparison of the sequences with other PRRSV isolates from around the world confirmed that the two ORFs were the ORF 5 (E gene) and ORF 6 (M gene) of the PRRSV genome. Sixteen overlapping base pairs were found between the coding region of E and M genes. The nucleotide homology with corresponding ORFs of the European PRRSV isolates (Lelystad, Boxmeer, PRRS-OLOT) varied from 52.1% to 53.7 % for E gene, and 63.8% to 64.8% for M gene. However, comparison of the Taiwanese M gene sequence with the American (VR2332, VR2385, 16244B, MN1), Canadian (IAF-Exp91), and Asian (EDRD-1 and CH-1a) isolates revealed very high degrees of homology (83.1% to 88.7% for E gene and 91.6% to 94.5% for M gene). Analysis of the deduced amino acid for E and M genes revealed that both proteins were very hydrophobic (42.5% for ORF5 and 43.7% for ORF6) which was consistent with their membrane-spanning character.

並列關鍵字

Porcine reproductive-end respiratory syndrome virus ； Membrane associated protein genes ； Sequence comparisons ； Taiwanese strain