[ Objective] The aim of this study was to clone and sequence F and HN genes of the isolated goose paramyxovirus and thus further investigate on its pathogenesis and evolutionary origins. [ Method] According to the pub...[ Objective] The aim of this study was to clone and sequence F and HN genes of the isolated goose paramyxovirus and thus further investigate on its pathogenesis and evolutionary origins. [ Method] According to the published F and HN gene sequences of Avian Paramyxovirus Typelat home and abroad, two pairs of primers （ FF, FR; HNF, HNR） were designed by DNAstar software, and Fand HNgenes were also amplified by PCR. The PCR products were recovered and ligated to T vector, while the positive clones were identified by ampicillin plate screening and PCR. After bacteria shaking, the plasmid was extracted and sent to Shanghai Sangon for sequencing. The sequences were compared with the sequences in GenBank, and the phylogenetic tree was drawn by DNAstar software. Meanwhile, the phylogenetic analysis of amino acid residues was also studied. [Result] The ORF of Fgene was 1 662 bp encoding 553 amino acids, and its cleavage site was 112R-R-Q-K-R-F117, which was consistent with the characteristics of virulent strains. 101-bit and 121-bit amino acid residues were K （Lys） and D （Asp）. The ORF of HN gene was 1 716 bp encoding 571 amino acids. The homology of HN sequence in 29 strains of goose paramyxovirus to YG97 was the highest, accounting for 99.6%. Analysis of glycosylation sites revealed that glycosylation site of BZ02 strain at 538 -540aa disappeared. The phylogenetic tree drawn by HN genes was highly consistent with that drawn by Fgenes. Compared with the homology of F and HN nucleotide sequence of the published typelgoose paramyxovirus in China, BZ02, JG97, HG97 and YG97 were divided into the same sub-group, and the nucleotide and amino acid sequences homologies were highest between BZ02 and YG97 strain. E Condusionl BZ02, JG97, HG97 and YG97 have the same evolutionary origin or all originate from YG97.展开更多
文摘[ Objective] The aim of this study was to clone and sequence F and HN genes of the isolated goose paramyxovirus and thus further investigate on its pathogenesis and evolutionary origins. [ Method] According to the published F and HN gene sequences of Avian Paramyxovirus Typelat home and abroad, two pairs of primers （ FF, FR; HNF, HNR） were designed by DNAstar software, and Fand HNgenes were also amplified by PCR. The PCR products were recovered and ligated to T vector, while the positive clones were identified by ampicillin plate screening and PCR. After bacteria shaking, the plasmid was extracted and sent to Shanghai Sangon for sequencing. The sequences were compared with the sequences in GenBank, and the phylogenetic tree was drawn by DNAstar software. Meanwhile, the phylogenetic analysis of amino acid residues was also studied. [Result] The ORF of Fgene was 1 662 bp encoding 553 amino acids, and its cleavage site was 112R-R-Q-K-R-F117, which was consistent with the characteristics of virulent strains. 101-bit and 121-bit amino acid residues were K （Lys） and D （Asp）. The ORF of HN gene was 1 716 bp encoding 571 amino acids. The homology of HN sequence in 29 strains of goose paramyxovirus to YG97 was the highest, accounting for 99.6%. Analysis of glycosylation sites revealed that glycosylation site of BZ02 strain at 538 -540aa disappeared. The phylogenetic tree drawn by HN genes was highly consistent with that drawn by Fgenes. Compared with the homology of F and HN nucleotide sequence of the published typelgoose paramyxovirus in China, BZ02, JG97, HG97 and YG97 were divided into the same sub-group, and the nucleotide and amino acid sequences homologies were highest between BZ02 and YG97 strain. E Condusionl BZ02, JG97, HG97 and YG97 have the same evolutionary origin or all originate from YG97.
