百合无症病毒衣壳蛋白基因克隆和蛋白分析

LSV coat protein genes cloning and protein analyses

根据已报道的LSV CP基因序列合成两条寡聚核苷酸引物,模板为感染LSV的百合叶片的总RNA,通过反转录-聚合酶链式反应(RT-PCR)扩增出大小为876bp的LSV CP基因,经测序后,对该基因编码区全长序列及相应的氨基酸序列用生物信息学软件系统进行序列分析及结构功能预测.结果表明:该基因由876个核苷酸组成,编码291个氨基酸;与GeneBank公布的其他LSV分离物的基因序列同源性为93.4%～99.0%,氨基酸同源性为84.8%～99.5%;它含有一个卷曲螺旋结构和多个磷酸化位点,平均疏水值为-0.432;含有Carlaviruses完整的衣壳蛋白保守结构域,二级结构以α-螺旋和无规则卷曲为主.

A pair of primers are designed and synthesized based on the oligosaccharides nucleotlde sequences of coat protein （CP） genes of lily symptomless virus （LSV） reported, then, a fragment of 876 bp of LSV CP gene sequence is amplified by reverse transcription polymerase chain reaction （RT- PCR） from the total RNA infected lily leaves. The function and the secondary structure of LSV CP are predicted by the bioinformatics software system after analyzing the sequence. The results show that the gene is 876 bp in length coding 291 amino acids. This isolation has a homology of 93.4^- 99.0~~ at nucleotide and 84. 8^-99. 5~ at amino acid levels respectively compared with those published sequences of LSV according to GeneBank. The functional study prediction shows that the grand average of hydropathicity is --0.432. The protein has one coiled-coil domain, several phosphorylation sites and a conserved structure domain of Carlaviruses CP. The secondary structure is mainly composed of a-helix and random coil-based structures.