关键词: 猪细小病毒; NS1基因; 克隆; 序列分析; 蛋白质结构预测

Abstract: A pair of specific primers was designed based on the sequence of NADL-2 strain and China strain of porcine parvovirus published in GenBank. The NS1 gene of PPV detected in Guizhou province was amplified, cloned, sequenced, sequence analyzed and protein structure predicted. The sequencing results showed that NS1 gene was 1986 bp in size,encoding 659 amino acids,the bases deletion of NS1 gene were found at site 1287,1288 and 1298;amino acids deletion were found at site 429,430 and 433. Phylogenetic tree showed that the NS1 gene sequencing in this experiment was at the same evolutionary branch with NADL-2 attenuated strain;the homologies of coding amino acids between this strain and NADL-2 attenuated strain and Kresse strain were both 98.6%. Protein structure prediction showed that molecular weight of non-structural protein NS1 was 75269.76 u, isoelectric pI was 7.25, instability coefficient was 41.57, speculating this protein was an instable protein. Aliphatic index was 73.58, the grand average of hydropathicity (GRAVY) was -0.565, suggesting that the protein was a hydrophilic protein. This protein contained rich α-helix,β-pleated sheet,β-turn and random coil, and there were many flexible regions, showing continuous distribution. No transmembrane region and signal peptide sequence were found in this protein;motif searching showed that non-structural protein NS1 might contain 3 N-glycosylation sites,3 cAMP- and cGMP-dependent protein kinase phosphorylation sites,5 protein kinase C phosphorylation sites,22 casein kinase Ⅱ phosphorylation sites,1 tyrosine kinase phosphorylation site and 8 N-myristoylation sites,1 amidation site and 1 ATP/GTP-binding site motif A (P-loop);10 major antigenic epitopes distributed in this protein.

Key words: porcine parvovirus; NS1 gene; cloning; sequence analysis; protein structure prediction