摘要
目的 了解人类博卡病毒2（HBoV2）基因突变对重要功能蛋白三级结够及其进化关系的影响.方法 对瑞安地区205份小儿腹泻患者粪便标本进行HBoV2基因筛选.选取阳性样本,采用巢式PCR方法对HBoV2全基因组扩增.通过Swiss-PdbViewer对预测蛋白结构进行分析,并采用MEGA5.1软件进行进化树分析.结果 瑞安地区HBoV2阳性检出率为5.36％ （11/205）.HBoV2全基因组经过拼接后为5196 bases,与参考基因组同源性达到99.79％.其中,NP1基因未发生突变,NS1、VP1和VP2基因存在有义突变.NS1测序组和对照组蛋白表面突变氨基酸疏水性质发生变化;VP1测序组和对照组蛋白空间结构和分子内部氢键分布发生变化.系统进化树分析显示,HBoV2中NSl突变造成亲缘关系变化,突变后NS1有向HBoV4演化趋势.结论 研究证明瑞安地区小儿腹泻患者存在HBoV2感染,且阳性样本中HBoV2基因发生突变.有义突变造成重要蛋白空间结构和亲缘关系变化.这些结果对研究HBoV2病毒突变后的进化方向奠定基础.
Objective To study the relationship among the genome mutation,viral protein structures and virus evolution of human Bocavirus 2（HBoV2）.Methods A method of PCR was applied to detect HBoV2 genome in 205 fecal specimens collected from children who had diarrhea in Ruian region,Zhejiang province,China.HBoV2-positive specimens were chosen to further amplify the HBoV2 genome.All the tertiary structures were predicted using the Swiss-PdbViewer software,and a phylogenetic analysis was performed by the MEGA software （version 5.1）.Result The positive rate of HBoV2 PCR detection in the collected 205 fecal specimens was 5.36% （11/205）.We obtained a sequence of a nearly full-length HBoV2 genome named as Ruian isolate,which is 5196 bases in length.Alignment of the Ruian HBoV2 sequence with other HBoV2 sequences in GenBank revealed that the new HBoV2 shared 99.79% similarities with the reference genome （GenBank no NC_012042）.No mutation was found in the NP1 gene,whereas sense mutation of NS1,VP1 and VP2 gene were found.Residue solvent accessibilities that relates to hydrophobic interactions were changed between the NS1 of the Ruian isolate and the reference isolate.The protein structure and location of H-bonds were different between the NS1 of the Ruian isolate and the reference isolate.The phylogenetic tree shows a clear division of the NS1,which has a potential evolution toward or a closely evolutionary relationship with the NS1 of HBoV4.Conclusion HBoV2 infection was detected in Ruian region,Zhejiang province.We found sense mutations in the HBoV2 genome of Ruian isolate.These mutations may alter HBoV2 protein structures,and may have an evolutionary significance.These findings provide data of HBoV2 genome mutation.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
2015年第2期133-135,共3页
Chinese Journal of Experimental and Clinical Virology
关键词
博卡病毒
人
腹泻
突变
蛋白质结构
三级
进化
Bocavirus,human
Diarrhea
Mutation
Protein structure,terciary
Evolution
