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Comparison of the nucleotide and deduced amino acid sequences of the S genes specified by virulent and avirulent strains of bovine coronaviruses

机译:牛冠状病毒强毒株和无毒株指定的S基因核苷酸和推导氨基酸序列的比较

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摘要

The entire nucleotide sequences of the spike glycoprotein (S) genes of the highly virulent bovine coronavirus (BCV) strain BCV-LY138, the avirulent BCV-L9 and related Norden Vaccine (BCV-Vaccine) strains were determined using the polymerase chain reaction (PCR) to amplify cDNAs obtained by reverse transcription of viral RNA, and to produce single strand cDNAs for DNA sequencing. The S gene sequences of these viral strains were compared with those of recently published strains BCV-Mebus, BCV-Quebec, and BCV-F15. An open reading frame of 4092 nucleotides, encoding a protein of 1363 amino acid residues, was found in all six strains. Frameshifts and insertions or deletions were not observed except for the BCV-Fl 5. The S gene sequences were more than 98% conserved overall inspite of different origins of the six viruses. There were 45 to 56 nt differences between the virulent and avirulent groups while there were 6 to 14 nt differences among four avirulent strains. Comparison of the deduced amino acid sequences indicated that the S proteins had typical properties of membrane glycoproteins. Nineteen Winked glycosylation sites were predicted in five strains, and 18 of them were conserved in the avirulent strain BCV-L9. The sequence KRRSRR at the predicted proteolytic cleavage site was identified in five strains while the sequence KRRSVR was found in BCV-Fl 5. Substitutions of few amino acids in the putative fusogenic domains and two prolines at 507 and 567 in the antigenic domains may cause altered immunogenic and other functional properties of the S proteins specified by the virulent and avirulent BCV strains. Nine amino acid substitutions between the virulent and avirulent groups may correlate with BCV virulence.
机译:使用聚合酶链反应(PCR)确定高毒力牛冠状病毒(BCV)菌株BCV-LY138,无毒BCV-L9和相关诺登疫苗(BCV-Vaccine)菌株的加标糖蛋白(S)基因的完整核苷酸序列。 )扩增通过病毒RNA逆转录获得的cDNA,并产生用于DNA测序的单链cDNA。将这些病毒株的S基因序列与最近发表的BCV-Mebus,BCV-Quebec和BCV-F15株的S基因序列进行了比较。在所有六个菌株中均发现了一个开放阅读框,编码4093个核苷酸,编码1363个氨基酸残基。除了BCV-F1 5外,未观察到移码,插入或缺失。尽管这六种病毒的起源不同,但S基因序列总体上保守了98%以上。毒力和无毒力组之间有45至56 nt的差异,而四种无毒力菌株之间有6至14 nt的差异。推导的氨基酸序列的比较表明,S蛋白具有膜糖蛋白的典型特性。在五个菌株中预测有19个眨眼糖基化位点,其中18个在无毒力BCV-L9菌株中保守。在五个菌株中鉴定出了在预测的蛋白水解切割位点处的序列KRRSRR,而在BCV-Fl 5中发现了序列KRRSVR。推定的融合结构域中的少数氨基酸被取代,而抗原结构域中的507和567处的两个脯氨酸被取代。由强毒和无毒BCV毒株确定的S蛋白的免疫原性和其他功能特性。有毒力和无毒力基团之间的九个氨基酸取代可能与BCV毒力相关。

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