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伪狂犬病病毒US1缺失株的构建和生物学特性分析
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作者 屈玮钰 李鑫 《中国兽医杂志》 北大核心 2025年第9期36-43,共8页
伪狂犬病病毒(PRV)是α疱疹病毒家族成员,其独特短区1(US1)基因在病毒生命周期中发挥关键作用,但US1在PRV中的功能研究尚不充分。本试验旨在构建PRV US1基因缺失株(PRV-ΔUS1),并探究其生物学特性。首先,通过蛋白表达纯化和动物免疫方... 伪狂犬病病毒(PRV)是α疱疹病毒家族成员,其独特短区1(US1)基因在病毒生命周期中发挥关键作用,但US1在PRV中的功能研究尚不充分。本试验旨在构建PRV US1基因缺失株(PRV-ΔUS1),并探究其生物学特性。首先,通过蛋白表达纯化和动物免疫方法制备鼠源PRV US1多克隆抗体,并利用成簇规律间隔短回文重复序列及其关联蛋白9(CRISPR/Cas9)技术构建PRV-ΔUS1,通过聚合酶链式反应(PCR)和蛋白质印迹法(WB)进行验证;然后,采用蚀斑试验、病毒生长曲线测定和连续传代遗传稳定性试验分析PRV-ΔUS1的生物学特性。结果显示,成功制备鼠源PRV US1多克隆抗体;经PCR和WB验证,PRV-ΔUS1成功缺失US1基因;与野生型毒株相比,PRV-ΔUS1的蚀斑直径极其显著缩小(P<0.001),6、12、24和48 h各个时间段的体外增殖能力均极其显著减弱(P<0.001),且在多次传代中保持遗传稳定性。结果表明,本试验成功构建PRV-ΔUS1,并证实US1基因可影响PRV的蚀斑形成和增殖能力,为PRV US1基因的功能研究和新型疫苗研发提供了重要工具和理论基础。 展开更多
关键词 伪狂犬病病毒(PRV) us1基因 CRISPR/Cas9 基因缺失病毒
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Characterization of Synonymous Codon Usage Bias in the Pseudorabies Virus US1 Gene 被引量:3
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作者 Meili Li Zhiyao Zhao +4 位作者 Jianhong Chen Bingyun Wang Zi Li Jian Li Mingsheng Cai 《Virologica Sinica》 SCIE CAS CSCD 2012年第5期303-315,共13页
In the present study, we examined the codon usage bias between pseudorabies virus (PRV) US1 gene and the USl-like genes of 20 reference alphaherpesviruses. Comparative analysis showed noticeable disparities of the s... In the present study, we examined the codon usage bias between pseudorabies virus (PRV) US1 gene and the USl-like genes of 20 reference alphaherpesviruses. Comparative analysis showed noticeable disparities of the synonymous codon usage bias in the 21 alphaherpesviruses, indicated by codon adaptation index, effective number of codons (ENc) and GC3s value. The codon usage pattern of PRV US1 gene was phylogenetically conserved and similar to that of the USl-like genes of the genus Varicellovirus of alphaherpesvirus, with a strong bias towards the codons with C and G at the third codon position. Cluster analysis of codon usage pattern of PRV US1 gene with its reference alphaherpesviruses demonstrated that the codon usage bias of USl-like genes of 21 alphaherpesviruses had a very close relation with their gene functions. ENc-plot revealed that the genetic heterogeneity in PRV US1 gene and the 20 reference alphaherpesviruses was constrained by G+C content, as well as the gene length. In addition, comparison of codon preferences in the US1 gene of PRV with those ofE. coli, yeast and human revealed that there were 50 codons showing distinct usage differences between PRV and yeast, 49 between PRV and human, but 48 between PRV and E. coil Although there were slightly fewer differences in codon usages between E.coli and PRV, the difference is unlikely to be statistically significant, and experimental studies are necessary to establish the most suitable expression system for PRV US1. In conclusion, these results may improve our understanding of the evolution, pathogenesis and functional studies of PRV, as well as contributing to the area of herpesvirus research or even studies with other viruses. 展开更多
关键词 Pseudorabies virus us1 gene ALPHAHERPESVIRUS Codon usage bias
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Cloning and sequence analysis of US1 gene in duck enteritis virus 被引量:1
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作者 ZHAO Yan WANG Jun-wei +1 位作者 MA Bo ZHAO Xiao-yan 《中国预防兽医学报》 CAS CSCD 北大核心 2011年第2期105-109,共5页
In this paper,a 1,860 bp sequence in IRs region of duck enteritis virus(DEV) was amplified by single oligonucleotide nested PCR with a single primer designed according to partial sequence of US1 and then a pair of pri... In this paper,a 1,860 bp sequence in IRs region of duck enteritis virus(DEV) was amplified by single oligonucleotide nested PCR with a single primer designed according to partial sequence of US1 and then a pair of primers designed according to the 3' UTR of US8 gene and 5' end of the new getting sequence were used to amplify a 2,426 bp sequence toward the TRs region.Sequence analysis revealed that the both sequences contained an identical 990 bp open reading frame of DEV US1 gene.The two ORFs were in opposite transcription orientation.Sequence comparison of the nucleotide sequence and the deduced amino acid sequence of US1 gene showed relatively high identity to Mardivirus.Phylogenetic tree analysis showed that the eleven herpesviruses viruses were classified into three groups,and the duck enteritis virus was most closely related to Mardivirus. 展开更多
关键词 摘要 编辑部 编辑工作 读者
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微机用户安全界面软件介绍
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作者 周健 乔永诚 《设计通讯》 1991年第1期43-44,共2页
关键词 微机 程序系统 us1
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