基于二代高通量测序技术的新型冠状肺炎病毒S朊突变点分析

Analysis of Mutation Sites of SRAR-CoV-2 Spike Glycoprotein Based on Second-Generation Sequencing

通过对美国华盛顿洲在2020年3至4月底爆发的新冠肺炎病人二代高通量测序数据分析,找出新冠病毒刺突糖蛋白(简称S朊)中存在的所有突变类型,为研究病毒在体内复制的突变规律及研究疫苗提供基础资料。利用NCBI中公布的130例美国华盛顿区报道的新冠肺炎病人二代高通量测序数据,进行序列组装,并对其朊编码基因进行深度突变分析,找出其潜在的疑似抗原变异位点(antigen variation,以下简称突变点)。排除30条未获全长的数据,共获得100份病人完整的SARS CoV 2序列,其S朊编码基因,主要突变点集中在S1区的SP区及受体结合区(RBD)之前的间隔区(突变区基本呈连续分布:126aa~153aa,194aa~204aa)和S2区的1250aa~1270aa区。100份样本的数据研究结果显示,新冠病毒S基因在病人体内复制过程中较为稳定,编码氨基酸的突变点(突变频率>15%)呈单样本散在分布,且S2区较S1区更为稳定。未在新冠病毒的RBD区找到突变率>20%的点,而S区散在零星突变区域主要集中在S1区间隔区(126aa~153aa,194aa~204aa处,且基本呈连续分布)和S2末端约20aa处。

All different mutation types existing in the spike glycoprotein(S protein)of coronavirus(SARS-CoV-2)were analyzed and searched out adopted second-generation high-throughput sequencing data,collected from March to the end of April 2020,from the outbreak of SARS-CoV-2 patients during March to the end of April 2020 in Washington state,US,to provide fundamental research data for studying on the mutation rules of SRAR-CoV-2 in vivo replication of the virus for studying on the vaccine.Second generation sequencing data of 130 cases with SAR-CoV-2 in Washington area US issued by NCBI were conducted sequences assemble,and conducted mutation analysis for their protein code genes in depth,to search out their latent suspected and/or possible antigen variation sites.A total of 100 patients′complete SRAR-CoV-2 sequences were obtained precluding 30 incomplete length data.The spike protein coded gene mainly concentrated on S1 domain and SP domain,as well as interval domain before receptor binding domain(RBD)(the mutation domain basically assumed as continuous distribution:126aa-153aa,194aa-204aa);1250aa-1270aa domain of S2 domain.Conclusion:the study on the results of data of 100 samples showed that the S gene of SRAR-CoV-2 patients′seemed stable during in vivo replication process,and the mutation sites of coded amino acids(mutation frequency>15%)and assumed as single sample diffused distribution,and S2 domain more stable than S1 domain.No mutation frequency>20%sites were searched out on RDB domain of SARS-CoV-2,while the diffused sporadic mutation domains mainly concentrated on the interval domain of S1 domain(126aa-153aa,194aa-204aa,and basically assumed as successive distribution)and at the end of S2 site 20 aa approximately.