禽腺病毒血清4型贵州株全基因序列分析

为了解禽腺病毒血清4型(FAdV-4)地方流行毒株的分子进化情况,基于实验室分离的2株FAdV-4贵州株GZ-BJ株和GZ-QL株,分别对2株FAdV-4毒株进行PCR分段扩增,扩增产物克隆至载体,提取质粒进行PCR和双酶切鉴定后筛选出重组质粒进行测序,将测序结果依次拼接得到病毒的全基因组,获得FAdV-4贵州株的全基因序列,并对其进行序列和遗传进化分析。结果显示,通过PCR分段扩增成功获得了2株FAdV-4贵州株(GZ-BJ株和GZ-QL株)的全基因序列,长度分别为43352、43723 bp,FAdV-4 GZ-BJ株全基因序列长度比FAdV-4 GZ-QL株短371 bp,少6个ORF(22K、putative 9.1 ku、u-exon、ORF17、ORF28、ORF42),二者的氨基酸同源性为57.1%。2株FAdV-4贵州株同国内外不同地区FAdV-4毒株核苷酸同源性在88.7%~100%,与FAdV-4经典毒株ON1比对,2株FAdV-4贵州株和国内FAdV-4分离株均缺失ORF19、ORF27、ORF30。系统进化树分析显示,2株FAdV-4贵州株GZ-BJ株和GZ-QL株仍属于Ⅰ群C种FAdV。研究结果表明,2株贵州株FAdV-4 GZ-BJ株和FAdV-4 GZ-QL株较国内外FAdV-4毒株均存在进化与突变,且FAdV-4 GZ-BJ株变化较大,但尚未改变其血清型,这为探索FAdV-4致病机理的分子机制研究提供依据。

Whole Genomic Sequencing Analysis of Fowl Adenovirus Serotype 4 from Guizhou

To comprehend the molecular evolution of fowl adenovirus serotype 4(FAdV-4)endemic strains,based on FAdV-4 GZ-BJ and FAdV-4 GZ-QL strains isolated in the laboratory,segmented amplification was performed on the two FAdV-4 strains,the amplified product was cloned into the vector,the plasmid was extracted,PCR and double enzyme digestion were performed,and the recombinant plasmid was screened for sequencing.The sequencing results were sequentially spliced to obtain the whole genome of FAdV-4 Guizhou strain,and sequence and genetic evolution analysis.The results showed that the lengths of two FAdV-4 Guizhou strains,GZ-BJ and GZ-QL strains,were 43352 and 43723 bp,were obtained by PCR fragmented amplification.respectively,and the complete genes of FAdV-4 GZ-BJ strain was 371 bp shorter than the FAdV-4 GZ-QL strain,and 6 ORFs(22K,putative 9.1 ku,u-exon,ORF17,ORF28,ORF42),and the amino acid homology of them was 57.1%.The nucleotide homology of the two FAdV-4 Guizhou strains with FAdV-4 strains in different regions at home and abroad were 88.7%-100%.The FAdV-4 two Guizhou strains and domestic isolates lacked ORF19,ORF27,ORF30 that compared with the FAdV-4 classic strain ON1.Phylogenetic tree analysis showed that the two Guizhou strains FAdV-4 GZ-BJ and FAdV-4 GZ-QL still belonged to groupⅠspecies C fowl adenovirus.This research indicated that the two Guizhou strains FAdV-4 GZ-BJ strain and FAdV-4 GZ-QL strain had evolution and mutations compared with domestic and foreign FAdV-4 strains,and FAdV-4 GZ-BJ strain had changed greatly,but not yet changing its serotype and provided a basis for exploring the molecular mechanism of FAdV-4 virus pathogenic mechanism.