科研動(dòng)態(tài)

為了解塞內(nèi)卡病毒分離株SVA/CH/ZZ/2016全基因組序列，設(shè)計(jì)4對(duì)相互重疊的特異性引物擴(kuò)增基因片段，將擴(kuò)增產(chǎn)物分別克隆至pCE2TA/Blunt-Zero載體并進(jìn)行測(cè)序，拼接校正后獲得SVA/CH/ZZ/2016株全基因組。結(jié)果顯示，該毒株基因組全長(zhǎng)7 292 bp，包括5'UTR（670 bp）、ORF（6 546 bp）以及3'UTR（76 bp）。選擇國內(nèi)外其他9株參考毒株序列，對(duì)編碼區(qū)12個(gè)基因的核苷酸及編碼氨基酸進(jìn)行比對(duì)。結(jié)果顯示，核苷酸同源性最高的是3B基因，最低的是VP1基因，其余基因的核苷酸序列同源性均在85.2%~100%之間。VP1基因遺傳進(jìn)化分析顯示，SVA/CH/ZZ/2016株與美國分離株USAIL_Purdue_43_2016和USAIN_Purdue_3698_2016株親緣關(guān)系最近，屬同一進(jìn)化分支，與原始毒株SVV-001株親緣關(guān)系最遠(yuǎn)。對(duì)SVA/CH/ZZ/2016株和原始毒株SVV-001 VP1蛋白的氨基酸序列進(jìn)行比對(duì)，發(fā)現(xiàn)共有10處氨基酸差異。本研究通過對(duì)SVA/CH/ZZ/2016株全基因組測(cè)序及分析，為進(jìn)一步開展SVA分子生物學(xué)研究及流行病學(xué)調(diào)查提供了基礎(chǔ)數(shù)據(jù)。

Sequencing and Analysis on Whole Genome of Senecavirus CH/ZZ/2016 Strain 

In order to identify the whole genome sequence of senecavirus SVA/CH/ZZ/2016 strain，four pairs of overlapping specific primers were designed to amplify gene fragments. The amplified products were cloned into pCE2TA/Blunt-Zero vector for sequencing. The whole genome of SVA/CH/ZZ/2016 strain was obtained after splicing and correction. The results showed that the total genome length of the strain was 7 292 bp，including 5'UTR（670 bp），ORF（6 546 bp）and 3'UTR（76 bp）. Other 9 reference strains both at home and abroad were selected and compared with the nucleotide and amino acid sequences of 12 genes in coding region. It was found that the nucleotide homology was highest in 3B gene，minimum in VP1 gene，and remained 85.2% to 100% in other genes. By phylogenetic analysis，it was shown that SVA/CH/ZZ/2016 strain had the closest genetic relationship with American isolates USAIL_Purdue_43_2016 and USAIN_Purdue_3698_2016，shared the same evolutionary branch，and was with largest evolutionary distance with the original strain SVV-001，a total of 10 amino acid differences were found. Therefore，the basic data was provided for further study on SVA molecular biology and epidemiological investigation.

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