水痘-带状疱疹病毒84-7株基因特征分析
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摘要
目的分析水痘-带状疱疹病毒(varicella-zoster virus,VZV)84-7株的基因特征。方法利用PCR法分别扩增VZV84-7株ORF38(PstⅠ)、ORF54(BglⅠ)、ORF62(SmaⅠ、Bss HⅡ、NaeⅠ)共5个酶切位点片段,运用生物信息学软件对5个酶切位点进行分析;PCR扩增病毒复制起点,分析VZV84-7株复制起点特征;PCR扩增VZV84-7株g E基因,对该基因进行序列分析。结果 VZV84-7株的酶切位点特征为:PstⅠ~+BglⅠ~+SmaⅠ~-Bss HⅡ~-NaeⅠ~-;复制起点区域包含1个13TA+19GA结构;g E基因序列与Oka株相比,在第1 170位存在1个碱基差异,为C→A,属于无义突变,但编码区氨基酸序列同源性为100%。结论 VZV84-7株的酶切位点和复制起点均有特异性,同时VZV84-7株与Oka株的g E基因仅有1个碱基突变,氨基酸序列同源性为100%。
Objective To analyze the genetic characteristics of varicella-zoster virus(VZV) 84-7 strain. Methods The ORF38(PstⅠ), ORF54(BglⅠ) and ORF62(SmaⅠ, Bss HⅡ and Nae Ⅰ) of VZV84-7 strain were amplified by PCR respectively, and analyzed for restriction site by bioinformatics software. The fragment of origin of virus replication was amplified by PCR and analyzed for charcteristics. The g E gene of VZV84-7 strain was amplified by PCR and sequenced.Results The restriction characteristrics of VZV84-7 strain was PstⅠ~+ BglⅠ~+ SmaⅠ~-Bss HⅡ~- NaeⅠ~-. The origin of virus replication included the structure of 13 TA + 19 GA differentiated from other strain of VZV. The g E gene sequence showed difference of C→A in one base at site 1 170 compared with that of Oka strain, which was a nonsense mutation. However,the homology of amino acid sequences in encoding regions of the two strains was 100%. Conclusion Both the restriction site and origin of replication of VZV84-7 strain were specific. Meanwhile, the g E gene of VZV84-7 strain showed only mutation in one base compared with that of Oka strain, while the homology of amino acid sequences of the two strains was100%.
Objective To analyze the genetic characteristics of varicella-zoster virus(VZV) 84-7 strain. Methods The ORF38(PstⅠ), ORF54(BglⅠ) and ORF62(SmaⅠ, Bss HⅡ and Nae Ⅰ) of VZV84-7 strain were amplified by PCR respectively, and analyzed for restriction site by bioinformatics software. The fragment of origin of virus replication was amplified by PCR and analyzed for charcteristics. The g E gene of VZV84-7 strain was amplified by PCR and sequenced.Results The restriction characteristrics of VZV84-7 strain was PstⅠ~+ BglⅠ~+ SmaⅠ~-Bss HⅡ~- NaeⅠ~-. The origin of virus replication included the structure of 13 TA + 19 GA differentiated from other strain of VZV. The g E gene sequence showed difference of C→A in one base at site 1 170 compared with that of Oka strain, which was a nonsense mutation. However,the homology of amino acid sequences in encoding regions of the two strains was 100%. Conclusion Both the restriction site and origin of replication of VZV84-7 strain were specific. Meanwhile, the g E gene of VZV84-7 strain showed only mutation in one base compared with that of Oka strain, while the homology of amino acid sequences of the two strains was100%.
引文
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[6]LIU J J.Genotyping and epidemiologic survey toward clinical varicella-zoster virus isolates collected in China[D].Anhui Medical University,2009.(in Chinese)刘静静.水痘-带状疱疹病毒临床分离株基因型别分析与流行病学调查[D].安微合肥:安徽医科大学,2009.
[7]FOWLER W J,GARCIA-VLACARCEL M,HILL-PERKINS M S,et al.Identification of immunodominant regions and linear B cell epitopes of the g E envelope protein of varicella-zoster virus[J].Virology,1995,214(2):531-540.
[2]LI X L,WANG X X,ZHANG Z Y,et al.Sequencing of whole genome of varicella-zoster virus 84-7 strain[J].Chin J Biologicals,2010,23(1):1-8.(in Chinese)李秀玲,王潇潇,张中洋,等.水痘-带状疱疹病毒VZV84-7株全基因序列测定及分析[J].中国生物制品学杂志,2010,23(1):1-8.
[3]LOPAREV V N,ARGAW T,KRAUSE P R,et al.Improved identification and differentiation of varicella-zoster virus(VZV)wild-type strains and an attenuated varicella vaccine strain using a VZV open reading frame 62-based PCR[J].J Clin Microbiol,2000,38(9):3156-3160.
[4]CAMPSALL P A,AU N H,PRENDIVILLE J S,et al.Detection and genotyping of varicella-zoster virus by Taq Man allelic discrimination real-time PCR[J].J Clin Micrbiol,2004,42(4):1409-1413.
[5]QUINLIVAN M,GERSHON A A,STEINBERG S P,et al.An evaluation of single nucleotide polymorphisms used to differentiate vaccine and wild type strains of varicella-zoster virus[J].J Med Virol,2005,75(1):174-180.
[6]LIU J J.Genotyping and epidemiologic survey toward clinical varicella-zoster virus isolates collected in China[D].Anhui Medical University,2009.(in Chinese)刘静静.水痘-带状疱疹病毒临床分离株基因型别分析与流行病学调查[D].安微合肥:安徽医科大学,2009.
[7]FOWLER W J,GARCIA-VLACARCEL M,HILL-PERKINS M S,et al.Identification of immunodominant regions and linear B cell epitopes of the g E envelope protein of varicella-zoster virus[J].Virology,1995,214(2):531-540.