Pudupakam Raghavendra Sumanth, Raghunath Shobana, Pudupakam Meghanath, Daggupati Sreenivasulu
Department of Veterinary Microbiology, College of Veterinary Science, Sri Venkateswara Veterinary University, Tirupati, Andhra Pradesh, India.
Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, USA.
Vet World. 2017 Mar;10(3):348-352. doi: 10.14202/vetworld.2017.348-352. Epub 2017 Mar 23.
Sequence analysis and phylogenetic studies based on non-structural protein-3 (NS3) gene are important in understanding the evolution and epidemiology of bluetongue virus (BTV). This study was aimed at characterizing the NS3 gene sequence of Indian BTV serotype-2 (BTV2) to elucidate its genetic relationship to global BTV isolates.
The NS3 gene of BTV2 was amplified from infected BHK-21 cell cultures, cloned and subjected to sequence analysis. The generated NS3 gene sequence was compared with the corresponding sequences of different BTV serotypes across the world, and a phylogenetic relationship was established.
The NS3 gene of BTV2 showed moderate levels of variability in comparison to different BTV serotypes, with nucleotide sequence identities ranging from 81% to 98%. The region showed high sequence homology of 93-99% at amino acid level with various BTV serotypes. The PPXY/PTAP late domain motifs, glycosylation sites, hydrophobic domains, and the amino acid residues critical for virus-host interactions were conserved in NS3 protein. Phylogenetic analysis revealed that BTV isolates segregate into four topotypes and that the Indian BTV2 in subclade IA is closely related to Asian and Australian origin strains.
Analysis of the NS3 gene indicated that Indian BTV2 isolate is closely related to strains from Asia and Australia, suggesting a common origin of infection. Although the pattern of evolution of BTV2 isolate is different from other global isolates, the deduced amino acid sequence of NS3 protein demonstrated high molecular stability.
基于非结构蛋白3(NS3)基因的序列分析和系统发育研究对于理解蓝舌病病毒(BTV)的进化和流行病学具有重要意义。本研究旨在对印度BTV血清型2(BTV2)的NS3基因序列进行特征分析,以阐明其与全球BTV分离株的遗传关系。
从感染的BHK - 21细胞培养物中扩增BTV2的NS3基因,进行克隆并进行序列分析。将生成的NS3基因序列与世界各地不同BTV血清型的相应序列进行比较,并建立系统发育关系。
与不同BTV血清型相比,BTV2的NS3基因显示出中等程度的变异性,核苷酸序列同一性范围为81%至98%。该区域在氨基酸水平上与各种BTV血清型具有93 - 99%的高序列同源性。PPXY/PTAP晚期结构域基序(motif)、糖基化位点、疏水域以及对病毒-宿主相互作用至关重要的氨基酸残基在NS3蛋白中是保守的。系统发育分析表明,BTV分离株分为四个拓扑型,亚分支IA中的印度BTV2与亚洲和澳大利亚起源的毒株密切相关。
NS3基因分析表明,印度BTV2分离株与亚洲和澳大利亚的毒株密切相关,表明存在共同的感染起源。尽管BTV2分离株的进化模式与其他全球分离株不同,但NS3蛋白的推导氨基酸序列显示出高分子稳定性。