马传染性贫血驴强毒gp90基因的克隆和序列分析

Cloning and sequencing of donkey-adapted equine infectious anemia virus gp90 gene

以EIAV驴强毒株D_AmRNA为模板 ,利用RT_PCR技术 ,扩增了约 1 .4kb的gp90基因。将其克隆后进行了测序 ,测序结果表明所扩增的 1 3 3 8个核苷酸片段含有完整的gp90基因全序列。核苷酸和氨基酸序列比较分析结果表明 :D_AEIAV与国内分离株辽系强毒L株差异率仅有 1 .8% ,而与国外毒株 (克隆 1 3 69,WENV1 7、WENV1 6、PSPEIAV1 9)核苷酸差异率在 3 5 .5 %～ 3 7.2 %之间 ;D_A株与国内分离株L株氨基酸水平差异率在 2 .9% ,而与国外毒株氨基酸水平上的差异率在 42 .6%～ 46.0 % ;D_AEIAV有 1 9个N_连接糖基化位点 ,L株、WENV1 7和WENV1 6是 1 8个 ,克隆 1 3 69、WENV1 6和WENV1 7亲本毒株PSPEIAV1 9是 1 2个。

Using reverse transcription_polymerase chain reaction technique, the gp90 gene of EIAV Chinese donkey_adapted strain(D_A EIAV) was amplified from the viral mRNA, and then cloned and sequenced. The sequenced result shows that the cloned cDNA fragment contains the whole opening reading frame of gp90 gene, which is 1 338 bp. Compared to other viruses, the gp90 has the highest homology with the isolate EIAV L strain at nucleotide and amino acid levels, the nucleotide diversity is only 1.8 % and has lower homology with foreign viruses(clone 1369, WENV17, WENV16 and PSPEIAV19),the nucleotide diversities were between 35.5 % and 37.2 %. The amino acid diversity was only 2.9 % between D_A strain and L strain, and the amino acid diversities were from 42.6 % to 46.0 % between D_A strain and foreign viruses. The D_A strain gp90 has 19 potential N_linked glycosylation sites, L strain, WENV17,and WENV 16 have 18 potential glycosylation sites, PSPEIAV19 and cloning 1369 have 12 glycosylation sites.