1株Ⅶ型新城疫病毒融合蛋白基因的克隆与生物信息学分析  

Cloning and Bioinformatics Analysis of FGene of Genotype Ⅶ Newcastle Disease Virus

作　　者：吴运谱[1,2] 赵晓彤[1,2] 薛树山[1,2] 石霖[1,2] 何利昆[1,2] 张晓华 曹明慧[4] 顾贵波[1,2] 谷志大[1,2] 魏澍[1,2] 曹东[1,2] 

机构地区：[1]辽宁省动物疫病预防控制中心,辽宁沈阳110164 [2]辽宁省动物医学研究院,辽宁沈阳110164 [3]辽宁省畜牧兽医局,辽宁沈阳110001 [4]中国兽医药品监察所,北京100081

出　　处：《中国畜牧兽医》2014年第7期68-73,共6页China Animal Husbandry & Veterinary Medicine

基　　金：辽宁省自然科学基金(201202099)

摘　　要：为了研究Ⅶ型新城疫病毒(Newcastle disease virus,NDV)CY1株的进化情况,分析其遗传变异和基因功能,试验应用RT-PCR扩增NDV辽宁分离株CY1株的融合蛋白(fusion,F)基因,进行测序分析,构建系统进化树,并用生物信息软件对蛋白结构功能进行分析。结果显示,NDVF基因全长1662bp,编码553个氨基酸。生物信息学分析结果表明,F蛋白的分子质量为58991.0u,理论等电点为8.48,半衰期为30h(哺乳动物类网状细胞),不稳定系数31.62,脂肪指数为108.12,总平均疏水性0.174,最大疏水性为3.078,最小疏水性为-2.578,信号肽序列为第1-31位氨基酸残基,跨膜区结构分析结果表明该蛋白为跨膜蛋白,预测可能含有6个N-糖基化作用位点、9个酪蛋白激酶Ⅱ磷酸化位点、7个蛋白激酶C磷酸化位点、14个N-豆蔻酰化位点、1个酰胺化位点、1个亮氨酸拉链、1个微体C端靶信号位点、1个双向核定位信号。系统进化分析结果表明,NDV CY1株与zj1株进化距离最近。The assay was aimed to analyze the protein structure and function of fusion （F） gene of genotype Newcastledisease virus （NDV） strain CY1. The total RNA was extracted from NDV CY1 strain, F gene was amplified by RT-PCR andsequenced, then using some related computer software and database, the sequence of F gene of NDV CY1 strain and the de-duced amino acid sequence were analyzed, and the secondary structure and hydrophobicity of the protein were determined andpredicted. The results showed that F gene of NDV CY1 strain was consisted of 1662 nucleotides encoding a 553-amino acidprotein. The protein had two transmembrane domains, and its subcellular localization was in plasma membrane and endoplasmicreticulum. The secondary structure contained several kinds of conformations. Grand average of hydropathicity of F protein ofNDV CY1 strain was 0. 174. The theoretical pI, estimated half-life, instability index and fatty index of F protein was 8.48,30 h （mammalian reticulocytes in vitro）, 31.62 and 108.12, respectively. The maximum and minimum hydropathicities were3. 078 and --2. 578 ,respectively. The protein included a 31-amino acid signal peptide. Motif search showed that F protein had 6N-glycosylation site, 9 casein kinase Ⅱ phosphorylation site, 7 protein kinase C phosphorylation site, 14 N-myristoylation site,1 amidation site, 1 leucine zipper pattern, 1 mierobodies C-terminal targeting signal, 1 bipartite nuclear localization signal pro-file. Analysis of phylogenetic tree indicated that the F gene of NDV CY1 strain had the highest genetic relationship with zjlstrain.

关 键 词：新城疫病毒 F基因 生物信息学分析 

分 类 号：Q78[生物学—分子生物学]