企鹅源禽1型副黏病毒基因组序列分析

Sequence Analysis of Complete Genome of Avian Paramyxovirus-1 Isolated from Penguin

为了从分子水平上探讨企鹅源禽1型副黏病毒BP01的演化,根据GenBank登录的鹅源禽1型副黏病毒基因序列设计引物,运用RT-PCR方法于国内外首次测定了企鹅源禽1型副黏病毒全基因组序列,其全长为15192nt。经分析表明BP01病毒基因组与SF02株和ZJ1株等鹅源禽1型副黏病毒同源性最高,分别为99.1%和99.0%;与IT-227/82和dove/Italy/2736/00等鸽源禽1型副黏病毒同源性分别为90.2%和88.1%;而与Hert/33和LaSota等鸡源禽1型副黏病毒的同源性最低,仅分别为87.9%和83.2%。对F基因第47～435nt进行系统进化树分析和F基因第334～1682nt进行HinfⅠ、BstOⅠ、RsaⅠ酶切位点分析,确定BP01株病毒为基因Ⅶ型。F蛋白裂解位点氨基酸序列为112R-R-Q-K-R-F117,符合禽1型副黏病毒强毒株的特性,经预测发现BP01株病毒F蛋白的七肽重复序列分别在第143～189位氨基酸残基和第461～503位氨基酸。通过对主要毒力基因推导的氨基酸序列进行比较发现,水禽源与其它源F蛋白有14个氨基酸不同,HN蛋白上有2个氨基酸的不同,而P蛋白有20个氨基酸的不同,且在第151～166位氨基酸存在一个明显的突变域。

To study the evolution of avian paramyxovirus-1 （APMV-1）BP01 strain isolated from penguin in molecular level,the whole genome of BP01 strain was amplified by RT-PCR based on primers designed according to paramyxovirus- 1 genomes published. Its complete genome was sequenced in the study,consisting of 15 192 nucleotide acids. This was the first reported at home and abroad. The higher identities up to 99.1% and 99.0% existed between BPO1 and APMVs-1 of SF02 and ZJ1 strains isolated from geese, separately. The homologies between BPO1 and APMVs-1 of IT-227/82 and dove/ ItalyF2736/00 isolated from pigeons were 90.2% and 88.1%, respectively. However, the lower identities between APMV-1 BP01 and APMVs-1 of Hert/33 and La Sota isolated from chickens were 87.9% and 83.2% ,separately. The F gene of BPO1 and published sequences were typing by phylogenetic tree （nt 47-435）and restriction enzyme cleavage site mapping （nt 334-1682）,revealing BPO1 strain belonged to genotype VII. The induced predicted amino acid sequence of cleavage site of fusion protein was ^112R-R-Q-K-R-F^117 and this result suggested that BP01 was velogenic. Prediction of the heptad repeat regions revealed that the heptad repeat regions located at two regions. The first region located at 143 to 189 amino acids of F protein,while the second at 461 to 503 amino acids. According to amino acid comparison of major virulent genes from waterfowl-origin APMVs-1 to that of other origin,there were 14,2 and 20 amino acids changed at F,HN and P protein separately. Especially,there was an evident mutation-motif located 151 to 166 of P protein of BPOI strain.