Cloning and further sequence analysis of the ORF3 gene of wild- and attenuated-type porcine epidemic diarrhea viruses.
Cloning and further sequence analysis of the ORF3 gene of wild- and attenuated-type porcine epidemic diarrhea viruses.
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DOI：
10.1007/s11262-007-0164-2
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发表时间：
2008-02
期刊：
影响因子：
1.6
通讯作者：
Park BK
中科院分区：
文献类型：
--
作者：
Park SJ;Moon HJ;Luo Y;Kim HK;Kim EM;Yang JS;Song DS;Kang BK;Lee CS;Park BK
为进一步研究猪流行性腹泻病毒(PEDV)的功能，对亲本DR13、减毒株DR13、KPED-9、P-5V和12个野毒株的ORF3基因进行了克隆和测序。测序结果表明，野生型PEDV ORF3基因的ORF为675个核苷酸，编码224个氨基酸的蛋白质，预测的相对分子质量为25.1-25.3 kDa。弱毒型PEDV ORF3基因的ORF为624个核苷酸，编码207个氨基酸的蛋白质，预测相对分子质量为23.4kDa。弱毒株包括减毒DR13、KPED-9和P-5V的ORF3基因编码区有51个核苷酸缺失，而野生型PEDV的ORF3基因包括CV777、BR1/87、LZC、亲本DR13和12个田间样品。此外，此前已发现减毒型PEDV在猪身上表现出较低的致病性。因此，51个核苷酸的缺失似乎是有意义的，可能对PEDV的致病性有重要意义，因为它们导致了减毒型PEDV预测的氨基酸序列的变化。对部分ORF3基因51个核苷酸缺失的逆转录聚合酶链式反应(RT-PCR)结果表明，所有PEDV都分为野生型和弱毒两种类型。含有亲本DR13和12个田间样品的野生型PEDV的RT-PCR产物为245bp，而含有PEDV疫苗株(减毒株DR13、KPED-9、P-5V)的减毒株PEDV的RT-PCR产物为194bp。此外，所有PEDV疫苗毒株都被用作活病毒疫苗，因为它们以前在猪身上表现出较低的致病性。因此，由51个核苷酸缺失导致的17个氨基酸缺失的大缺失区可能是PEDV致病的重要部位，可用于区分野生型和弱毒型。
The open reading frame (ORF3) genes of the parent DR13, attenuated DR13, KPED-9, P-5V, and 12 field samples were cloned and sequenced to further explore the functions of wild- and attenuated-type porcine epidemic diarrhea viruses (PEDVs). Sequencing revealed that wild-type PEDVs ORF3 genes had a single ORF of 675 nucleotides encoding a protein of 224 amino acids with a predicted M r of 25.1–25.3 kDa. Attenuated-type PEDVs ORF3 genes had a single ORF of 624 nucleotides encoding a protein of 207 amino acids with a predicted M r of 23.4 kDa. The coding region of the ORF3 gene of attenuated-type PEDVs including attenuated DR13, KPED-9, and P-5V had 51 nucleotide deletions that were not found in the ORF3 genes of wild-type PEDVs including CV777, Br1/87, LZC, parent DR13, and 12 field samples. In addition, attenuated-type PEDVs have previously been found to exhibit reduced pathogenicity in pigs. Therefore, 51 nucleotide deletions appear to be meaningful and may be significant for PEDV pathogenicity, because they lead to changes in the predicted amino acid sequences of attenuated-type PEDVs. Reverse transcriptase-polymerase chain reaction (RT-PCR) on the partial ORF3 gene including 51 nucleotide deletions revealed that all PEDVs fell into two types, wild- and attenuated-type PEDVs. Wild-type PEDVs containing parent DR13 and 12 field samples had RT-PCR products of 245 bp in size, while attenuated-type PEDVs containing PEDV vaccine strains (attenuated DR13, KPED-9, P-5V) had products of 194 bp. In addition, all PEDV vaccine strains were used as live virus vaccine, because they previously exhibited a reduced pathogenicity in pigs. Therefore, large deletion region, which is comprise 17 amino acid deletions caused by 51 nucleotide deletions and is seen in all PED live vaccine strains, may be important site for PEDV pathogenicity, and we can use it for differentiation of wild- and attenuated-type PEDVs.
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影响因子：
1.2
作者：
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通讯作者：
Jackwood, DJ
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影响因子：
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作者：
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