Cloning and further sequence analysis of the spike gene of attenuated porcine epidemic diarrhea virus DR13.
Cloning and further sequence analysis of the spike gene of attenuated porcine epidemic diarrhea virus DR13.
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肿瘤病毒腹泻病毒DR13的尖峰基因的克隆和进一步的序列分析。
DOI：
10.1007/s11262-006-0036-1
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发表时间：
2007-08
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;Song DS;Ha GW;Park BK
为进一步研究猪流行性腹泻病毒（PEDV）弱毒和野生型PEDV的功能，克隆了猪流行性腹泻病毒（PEDV）DR 13株S基因。测序结果表明，该基因编码一个4,149个核苷酸的开放阅读框，编码1,382个氨基酸，分子量为151 kDa。减毒PEDV DR 13的S基因的编码区具有20个核苷酸变化，这些变化似乎是功能的重要决定因素，因为它们在其预测的氨基酸序列中产生变化。值得注意的是，先前已经发现减毒PEDV DR 13在猪中表现出降低的致病性。因此，含有这20个核苷酸变化的区域可能对PEDV致病性至关重要。减毒的PEDV DR 13 S蛋白含有28个Asn-Xaa-Ser/Thr序列子、21个预测为N-糖基化的天冬酰胺和在位置1，327 - 1，347处的一段高度疏水残基，预测其形成α-螺旋并用作膜锚。推导的氨基酸序列中的一个变化（从378处的N到K）破坏了N-连接的糖基化位点，而另一个变化（从114处的N到S）在不同的位置产生了一个新的糖基化位点。这些N-连接糖基化位点的改变反映了3个核苷酸的变化，这些变化与上述核苷酸变化有关，并建议影响减毒PEDV DR 13的致病性。与CV 777、Br 1/87、JS-2004-2、Spk 1、Chinju 99和亲本DR 13的DNA序列同源性分别为96.5%、96.4%、96.1%、93.9%、93.5%和96.6%。在推导的氨基酸序列水平上，与这些基因的同源性分别为95.7%、95.4%、95.6%、92.0%、91.6%和95.7%。系统进化分析表明，PEDV减毒株DR 13与CV 777、Br 1/87、JS-2004-2和亲本DR 13的亲缘关系较近，与Spk 1和Chinju 99的亲缘关系较远，与中国PEDV JS-2004-2的亲缘关系较近。
The spike (S) gene of the attenuated porcine epidemic diarrhea virus (PEDV) DR13 was cloned and sequenced to further explore the functions of wild type PEDV and attenuated PEDV. Sequencing revealed a single large ORF of 4,149 nucleotides encoding a protein of 1,382 amino acids with predicted M r of 151 kDa. The coding region of the S gene of attenuated PEDV DR13 had 20 nucleotide changes that appeared to be significant determinants of function in that they produced changes in its predicted amino acid sequence. Notably, attenuated PEDV DR13 has previously been found to exhibit reduced pathogenicity in pigs. The regions containing these 20 nucleotide changes may therefore be crucial for PEDV pathogenicity. The attenuated PEDV DR13 S protein contains 28 Asn-Xaa-Ser/Thr sequons, 21 asparagines that are predicted to be N-glycosylated and a stretch of highly hydrophobic residues at positions 1,327–1,347, which is predicted to form an α-helix and to function as a membrane anchor. One (from N to K at 378) of the changes in the deduced amino acid sequence destroyed N-linked glycosylation sites, while another change (from N to S at 114) created a new one at a different location. These alterations in N-linked glycosylation sites reflected 3 nucleotide changes, which were related to the above-mentioned nucleotide changes and are suggested to influence the pathogenicity of attenuated PEDV DR13. Attenuated PEDV DR13 has 96.5, 96.4, 96.1, 93.9, 93.5 and 96.6% DNA sequence identities with CV777, Br1/87, JS-2004-2, Spk1, Chinju99 and parent DR13, respectively. Likewise, it shares 95.7, 95.4, 95.6, 92.0, 91.6 and 95.7% identity with those genes at the deduced amino acid sequence level. Phylogenetic analysis suggested that attenuated PEDV DR13 is closely related to CV777, Br1/87, JS-2004-2 and parent DR13, rather than to Spk1 and Chinju99 and is especially close to the Chinese PEDV strain JS-2004-2.
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