【摘要】 本研究应用11日龄SPF鸡胚增殖禽流感病毒标准株A/Duck/Germany/1215(H2N3),用LSTRIAZOL试剂盒提取病毒RNA,应用自行设计的NA基因半特异性引物进行RT＿PCR扩增,得到NA基因的全长DNA片段,克隆到PMD18＿T载体上,并进行鉴定和序列测定。测定结果表明所获得的NA基因DNA片段长1451bp,编码469个氨基酸残基,与其他亚型的NA基因编码的氨基酸长度相一致。根据推导的氨基酸序列进行预测,该蛋白具有6个潜在的糖基化位点和19个半胱氨酸残基。与其他亚型的NA基因相比较发现克隆的NA基因符合神经氨酸的分子结构特征,本研究首次获得N3亚型NA基因全序列。更多还原

【Abstract】 A N3 subtype influenza virus A/Duck/Germany/1215/73(H2N3) was grown in 11-day-old SPF embryonated chicken eggs.Virial RNA was isolated by treatment of aliquots of the virus with LS TRIAZOL.Full-length NA cDNA was prepared by RT-PCR and cloned into pMDT-18 vector.Sequences proved that the cloned NA gene consist of 1495 nucleotides and encode a polypeptide of 469 amino acids residues.The N3 NA protein has 6 potential glycosylation site and 19 cysteines.The cloned gene has the same molecular character of influenza NA gene compare to the other subtypes neuraminidase.更多还原