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[中国新疆绵羊肺腺瘤病毒的病理诊断及全基因组序列分析]

[Pathological Diagnoses and Whole-genome Sequence Analyses of the Jaagsiekte Sheep Retrovirus in Xinjiang, China].

作者信息

Yang Sufang, Liang Tian, Zhao Qingliang, Zhang Dianqing, Zhang Jing, Yang Xia, Sheng Jinliang

出版信息

Bing Du Xue Bao. 2015 May;31(3):217-25.

Abstract

To carry out pathologic diagnoses and whole-genome sequence analyses of the Jaagsiekte sheep retrovirus (JSRV) in Xinjiang, China, we first observed sheep suspected to have the JSRV. Then, the extracted virus suspension was observed by transmission electron microscopy (TEM). Total RNAs from lungs of JSRV-infected sheep were extracted and reverse-transcribed using a cDNA synthesis kit. Six pairs of primers were designed according to the exogenous reference virus strain (AF105220). Reverse transcription-polymerase chain reaction was carried out from JSRV-infected tissue, and the whole genome of the JSRV sequenced. Our results showed: flow of nasal fluid ("wheelbarrow test"); different sizes of adenoma lesions in the lungs; papillary hyperplasia of alveolar epithelial cells; alveolar cavity filled with macrophages; dissolute nuclei in central lesions. TEM revealed JSRV particles with a diameter of 88 nm to 125. 4 nm. The full-length of the viral genome sequence was 7456 bp. BLAST analyses showed nucleotide homology of 96% and 95% compared with that of the representative strain from the USA (AF105220) and UK (AF357971). Nucleotide homology was 89.8% and 89.9% compared with the endogenous Jaagsiekte sheep retrovirus, Inner Mongolia strain (DQ838493) and USA strain (EF680300). The specific pathogenic amino-acid sequence "YXXM" was found in the TM district, similar to the exogenous JSRV: this gene has been reported to be oncogenic. This is the first report of the complete genomic sequence of the exogenous JSRV from Xinjiang, and could lay the foundation for study of the biological characteristics and pathogenic mechanisms of the pulmonary adenomatosis virus in sheep.

摘要

为对中国新疆绵羊肺腺瘤病毒(JSRV)进行病理诊断及全基因组序列分析,我们首先对疑似感染JSRV的绵羊进行观察。然后,利用透射电子显微镜(TEM)观察提取的病毒悬液。从感染JSRV的绵羊肺组织中提取总RNA,并使用cDNA合成试剂盒进行逆转录。根据外源性参考病毒株(AF105220)设计了6对引物。从感染JSRV的组织中进行逆转录-聚合酶链反应,并对JSRV的全基因组进行测序。我们的结果显示:鼻液流出(“手推车试验”);肺内大小不同的腺瘤病变;肺泡上皮细胞乳头样增生;肺泡腔内充满巨噬细胞;中央病变区细胞核溶解。TEM显示JSRV颗粒直径为88 nm至125.4 nm。病毒基因组序列全长7456 bp。BLAST分析显示,与美国代表性毒株(AF105220)和英国代表性毒株(AF357971)相比,核苷酸同源性分别为96%和95%。与内源性绵羊肺腺瘤病毒内蒙古株(DQ838493)和美国株(EF680300)相比,核苷酸同源性分别为89.8%和89.9%。在跨膜区发现了特异性致病氨基酸序列“YXXM”,与外源性JSRV相似:该基因已被报道具有致癌性。这是新疆外源性JSRV完整基因组序列的首次报道,可为绵羊肺腺瘤病病毒生物学特性及致病机制的研究奠定基础。

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