摘要
将古浪山羊痘病毒的基因组DNA提取出来,并设计引物进行PCR扩增,克隆F10L基因的DNA序列.为了分析山羊痘病毒F10蛋白的分子特征,将PCR产物连接到p GEM-T Easy载体后转化至大肠杆菌DH5α感受态细胞,筛选阳性克隆进行序列测定,利用生物信息学软件对F10L基因序列进行预测分析.结果显示,F10L基因序列由1 335个核苷酸组成的开放阅读框,编码444个氨基酸残基组成的多肽,蛋白质分子质量的理论值53.01 ku,理论等电点为7.14.该蛋白的二级结构中,α螺旋占12.44%,β折叠占15.73%,其余71.83%为无规则卷曲.多序列比对分析显示,不同羊痘病毒分离株F10L序列高度保守,一致性在97%以上.此研究结果为进一步研究F10蛋白的生物学功能和羊痘病毒早期蛋白的分子相互作用奠定了基础.
This study was extracted genomic DNA from Goatpox virus GuLang isolate. The specific primers were designed and used to amplify the F10L gene from the genomic DNA by PCR. A PCR product was ligated into pGEM - T Easy vector. After transformation into E coli DH5a, the positive clones were sequenced and the sequences were analyzed with the bioinformatic softwares. To explore the molecular characteristics of F10L from Goatpox virus（GTPV）. The result showed F10L gene sequence contained an open reading frame（ORF） of 1 335 nucleotides and deduced protein consisted of 444 amino acids with the theoretical molecular weight of 52.01 Ku and isoelectxic point of 7.14. Analysis of secondary structure of F10 revealed that a- helix, [3 - strand and loop were 12.44 %, 15.73 % and 71.83 %, respectively. Analysis of multiple sequence alignment indicated F10L gene from different Capripoxvirus isolates were highly conserved with identity of 97 %, The present results laid the foundation for further studies of biological functions of F10 and interaction among the early proteins of Capripoxvirus.
出处
《西北民族大学学报（自然科学版）》
2014年第4期69-74,共6页
Journal of Northwest Minzu University(Natural Science)
基金
国家质检总局公益性行业科研专项(201310093)
国家自然基金项目(31201892)
甘肃省自然基金项目(1208RJZA101)
国家863计划项目(2012AA101304)资助
