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1.
中国狂犬病毒设苗株(5aG株)糖基因的克隆与序列分析 总被引:8,自引:0,他引:8
用逆转录-聚合酶链反应方法从中国犬疫苗株病毒感染细胞中扩增得到该株病毒糖蛋白基因,并进行序列测定。结果表明该基因开放阅读框架全长1575bp,编码505个氨基酸的成熟糖N-端19个氨基酸的信号肽,该基因与其他株系相应基因比较,核酸序列同源性为88%-91%,氨基酸序列同源性为87-90%,其中膜外区同源性高于膜内区及跨膜区同源性。 相似文献
2.
本文报告了中国广西狂犬病毒野毒株(CGX89-1株)糖蛋白基因cDNA的核苷酸序列及其推导的氨基酸序列。CGX89-1株的糖蛋白基因从起始密码ATG到终止密码TAA共有1575个核苷酸残基,可编码形成524个氨基酸残基的多肽链,经修饰后形成具有505个氨基酸残基构成的狂犬病毒糖蛋白。CGX89-1株的糖蛋白基因和核苷酸组成分别为:A占27.11%,T占26.29%,C占21.97%和G占24.63%。核苷酸序列和巴斯德株(PV株),国际标准攻击毒株(CVS株),中国狂犬病毒疫苗株(3aG株)相比,同源性分别为84.1%,83.1%和84.5%。其推导的氨基酸序列和PV株、CVS株和3aG株相比其同源性分别为92.4%,89.7%和89.5%。CGX89-1株也具有3个N-糖基化位点,分别位于第37位、157位和319位的氨基酸残基上。糖蛋白的膜外区部分重要抗原位点和PV株、CVS株及3aG株具有较高的一致性。 相似文献
3.
狂犬病毒5aG株核蛋白基因的克隆及序列分析 总被引:8,自引:0,他引:8
用RT、RACE、PCR等方法,从中国狂犬疫苗株(5aG)病毒感染的鼠脑组织中扩增病毒核蛋白基因,用双链测序法进行了全基因核酸序列分析。结果表明:该基因开放阅读框架全长1353bp,编码450个氨基酸。与其它3株狂犬病毒核蛋白相比,核酸序列同源性为90%~96%,氨基酸序列同源性为95%~96%。表明核蛋白保守性较好。 相似文献
4.
5株呼吸道合胞病毒地方株F蛋白基因序列分析 总被引:2,自引:0,他引:2
目的呼吸道合胞病毒(RSV)F蛋白是RSV感染免疫中最重要的病毒蛋白,为了解我国RSV地方株F蛋白的基因状况和变异特征,随机选取北京、广州、长春和河北四个地区具有不同流行特征的RSV地方株(A亚型)5株,进行RSVF蛋白全基因的核苷酸序列分析。方法以提取的病毒mRNA为模板进行RT-PCR扩增、目的基因的克隆及序列测定,对地方株及原型株的序列进行比较分析。结果地方株F蛋白基因与原型株A2株有很高的同源性,核苷酸全序列的同源性为95.1%~96.1%,氨基酸同源性为96.7%~97.4%。核苷酸有义突变率为22.6%~25.9%。3非编码区的核苷酸序列比蛋白编码区变异显著。河北地方株(E73株)在3非编码区有6个核苷酸的插入。F2亚单位的氨基酸变异高于F1亚单位。在北京地方株(ZHS13株)F1亚单位内,由具有中和能力单克隆抗体所识别的抗原表位区中存在一个氨基酸的变异。结论我国RSV地方株与原型株之间的F蛋白基因尽管存在一定的变异,但仍有很高的同源性。地方株间F蛋白的核苷酸、氨基酸变异的位置及形式很相似,提示我国RSV的不同流行特征可能并非由于F蛋白的基因变异所致。 相似文献
5.
汉滩病毒Z10株G1糖蛋白编码区克隆及序列分析 总被引:1,自引:0,他引:1
目的 汉滩病毒浙10(Z10)株G1糖蛋白编码区的克隆、核苷酸序列分析,提供我国应用最为广泛的肾综合征出血热(HFRS)疫苗株序列资料。方法 反转录PCR法扩增G1基因片段,克隆入pGEM-T载体,双脱氧链终止法测定核苷酸序列。结果 测定1449个核苷酸,可编码483个氨基酸。与I型76/118、Lee、Hojo株比较核苷酸同源性分别为87%、86%、86%,与Ⅱ型R22株同源性为67%。比较氨基 相似文献
6.
中国分离乙脑病毒与灭活疫苗株(P3株)E基因差异分析 总被引:5,自引:1,他引:5
目的 分析我国近年来从蚊虫及患者标本中分离的乙脑病毒与灭活疫苗株(P3株)之间在E基因区段核苷酸及氨基酸差异。方法 从GenBank中获取相应乙脑病毒株E基因区段核苷酸序列,通过Clustal X(1.8)、DNASTAR、GENEDOC(3.2)等生物学软件进行分析。结果 P3株与福建分离株之间核苷酸同源性在98.3%-98.5%之间、氨基酸同源性在98.2%-98.6%之间;P3株与上海分离株之间核苷酸同源性在88.0%-88.5%之间、氨基酸同源性在98.0%-98.4%之间。E基因区段500个氨基酸中P3株与所有新分离乙脑病毒株之间共存在19个位点的差异,其中在E-76、E-306、E-408处所有新分离毒株与P3株存在共同的差异;在E-160、E-487处福建分离株与P3株存在共同差异;上海分离株与P3株在E-129、E-222、E-227、E-366存在共同差异。结论 上海蚊虫中分离的Ⅰ型乙脑病毒和福建省脑炎患者中分离的Ⅲ型乙脑病毒与P3株在E基因的部分氨基酸位点存在差异,但均不处在影响病毒生物学特性的关键位点。 相似文献
7.
本文报道1987年从广西病人血清中分离的登革2型(D2)病毒D2-43株和1985年从海南病人血清中分离的D2-04株乳鼠致病性的差异与基因变化的关系。结果表明D2-43株对乳鼠致病,D2-04株不致病。D2-43株和D2-04株C到NS1基因的读码框架基本相同,均由3381核苷酸组成,编码氨基酸总数1127。包含三个结构蛋白C.PrM(M)、E和一个非结构蛋白NS1。该两株病毒核苷酸序列同源性为93.8%,氨基酸序列的类似性为91.3%。C和E基因同源性为95.0%~95.8%,NS1基因为92.2%,M基因为86.7%,两株之间核苷酸序列的主要差异是在M基因。两株病毒的C-NS1基因与国际参考毒株比较,43株与JAM株类侧性最高,其次是NGC株,与S1株类似性最小。而04株只有C、E和NS1与JAM株最高,PrM(M)都与NGC株最高。43株与04株的M基因相比较,04株的PrM(M)基因的同源性低于43株,说明两株与国际参考毒株比较,主要差异也在M基因,乳鼠致病性差异可能与M基因变化有关。 相似文献
8.
痘苗病毒天坛株基因组病毒生长因子基因结构的研究 总被引:2,自引:0,他引:2
克隆了位于我国痘苗病毒天坛株基因组左端的病毒生长因子(VGF)基因并对其编码多肽的结构特点进行了分析。结果表明,由天坛株VGF基因所推导的氨基酸序列具有典型的上皮生长因子(EGF)超家族成员的特征序列和结构特点,在核苷酸及氨基酸水平上天坛株VGF与其他正痘病毒VGF的同源性在85%~95%之间,氨基酸残基的缺失和插入等变异主要集中出现在多肽的信号肽和穿膜区,推测对功能的影响不大。此外,我们发现由天坛株基因组右末端的开放读码框架(ORF)TB22L推导的氨基酸序列与天坛株VGF多肽第67位至140位完全相同,提示ORFTB22L可能是天坛株基因组第二个VGF基因的变异产物。 相似文献
9.
中国广西狂犬病毒野毒株(CGX89—1株)糖蛋白基因核… 总被引:3,自引:1,他引:3
本报告了中国广西狂犬病毒野毒株(CGX89-1株)糖蛋白基因cDNA的核苷酸序列及其推导的氨基酸序列。CGX89-1株的糖蛋白基因从起始密码ATG到终止密码TAA共有1575个核苷酸残基,可编码形成524个氨基酸残基的多肽链,经修饰后形成具有505个氨基酸残基构成的狂犬病毒糖蛋白。CGX89-1株的糖蛋白基因和核苷酸组成分别为:A占27.11%,T占26.29%,C占21.97%和G占24.63% 相似文献
10.
利用反转录多聚酶链反应(RT-PCR)技术分离并克隆了登革4型01株病毒的包膜蛋白(E)基因,并测定了部分核苷酸序列,发现该株与国外发表的一株相应区域核苷酸序列的同源性为93.4%,由此推测出氨基酸序列的同源性为92.1%。 相似文献
11.
Rabies virus (RV) glycoprotein (gp) consists of three domains: cytoplasmic, transmembrane and ectodomain. It occurs in a complete, membrane-bound form within the infected cell, but it is released from them in a deleted, secreted form lacking the transmembrane domain. This study was performed to test the importance of the transmembrane domain for the capability of the RV gp gene, introduced into mice via a recombinant plasmid (DNA vaccine), to induce immune response and protection against challenge. Although the antibody response to the secreted form of gp was higher than that to complete gp, the protective efficacy of the respective DNA vaccine against challenge was not better than that of the DNA vaccine inducing complete gp. This indicates that the transmembrane domain of RV gp is important for generating protection against rabies and should be present in RV DNA vaccines. 相似文献
12.
Summary The glycoprotein gene of the rabies virus vaccine strain Vnukovo-32 was sequenced and the deduced protein sequence was analyzed and compared with that of various laboratory and street strains. The amino acid sequence homologies of strain Vnukovo-32 were compared with fixed strains ERA, SAD B19, PV, HEP-Flury, CVS and two street strains, canine and CXX89-1, were 98.9 %(6 replacements), 98.3% (9), 96.2% (20), 91.4% (45), 87.0% (68), 93.5% (34) and 91.4% (45), respectively. Sequence alignments of the proteins revealed that the most conserved region is the ectodomain, whereas the transmembrane and cytoplasmic domains showed significant divergence. 相似文献
13.
Rabies virus glycoprotein (G) is a trimeric type I transmembrane glycoprotein that mediates both receptor recognition and low pH-induced membrane fusion. Electron microscopy has indicated that the ectodomain of protein G is made of a globular head and a stem. In order to characterize the putative stem region at the molecular level, we designed two peptides, P(S) and P(L), which were produced as GST fusion proteins in bacteria. Peptide P(S) extends from amino acid (aa) 374 to aa 428 whereas peptide P(L) extends from aa 368 down to the end of the ectodomain of G (aa 439). Their secondary and quaternary structures have been studied with spectroscopic and biophysical methods. We show that these isolated peptides are monomeric and poorly structured in aqueous solution. However, circular dichroism (CD) in presence of 2,2,2-trifluoroethanol and NMR data indicate that this region may adopt a alpha-helical conformation in the complete glycoprotein. 相似文献
14.
Rabies is an important public health problem in South East Asia, with cases in this part of the world contributing to about
70% of the global burden. A large number of rabies cases occur in India, however, there is no organized system of surveillance
and hence there is a lack of reliable data. Moreover, comprehensive molecular epidemiological studies have not been performed
on Indian virus isolates. In this study, we determined the complete nucleotide and deduced amino acid sequence of a primary
isolate of rabies virus obtained from the brain of an infected patient. Comparison of the genomic sequence with those of the
ten fully sequenced rabies strains available in GenBank showed nucleotide homology ranging from 97% with AY956319 to 81% with
AY705373. Amino acid homology of nucleoprotein ranged from 99.7% with AY352493 to 92% with DQ875051. In case of the glycoprotein
gene, the homology ranged from 98.8% with AY956319 to 87.2 % with AY705373. An extensive nucleoprotein, glycoprotein, and
full-length genome-based phylogenetic analysis was performed along with sequences available from the GenBank. Phylogenetic
analysis of the complete genome sequence indicated that this isolate exhibited close homology with the ex Indian strain AY956319.
Primer sequences and the scheme of amplification can be availed from the authors on request. 相似文献
15.
中国狂犬病毒疫苗株(5aG)糖蛋白基因在痘苗病毒中的表达及免疫效果观察 总被引:1,自引:0,他引:1
将克隆到的中国狂犬病毒疫苗株(5aG)的糖蛋白基因重组到痘苗病毒TK区,并在痘苗病毒P11启动子的控制下,构建了狂犬-痘苗重组病毒(VVaG)。经间接免疫荧光和Western免疫印染证明,重组病毒VVaG能良好地表达狂犬病毒糖蛋白,其分子量约为6600。用VVaG免疫小鼠,7d便可诱生较高的狂犬病毒中和抗体,21d达4169,并能100%保护狂犬病毒本毒株和国际标准攻击毒(CVS)的致死量攻击。 相似文献
16.
Rabies occurs in all parts of Indian sub-continent except Andaman and Nicobar and Lakshadweep group of islands. The full-length nucleoprotein (N) gene sequence of a rabies virus isolate from India is reported for the first time and the same has been compared with available N gene sequences from the database. A central domain of 230 amino acids (aa) from aa 141 to aa 370 exhibited more than 95% similarity. There were 8 amino acid positions (aa 29, 32, 38, 84, 119, 379, 438, and 439) at which substitution was unique for Indian isolates but common for laboratory strains. In antigenic epitopes, except for a single amino acid difference at the antigenic site IV, the amino acids were conserved. The Indian isolate also possessed two Bam HI sites (aa 247 and 278), while the other Asian isolates had only one site at aa 278 or were not digested with Bam HI at all. Phylogenetic analysis also demonstrated that the Indian isolate was closely related to the Sri Lankan isolate and grouped in the cluster that comprised of the isolates from other Asian countries namely China and Pakistan. 相似文献
17.
The gene encoding the attachment glycoprotein (G) was sequenced in three French isolates of-subgroup C avian metapneumovirus (APV-C) from ducks. With 1771 nt, this gene proved as long as recently published for North-American APV-C isolates from turkeys. The nt sequences of the duck viruses shared 99% identity but proved only 75-83% identical with their North-American counterparts, viruses of both origins encoding 585 amino acid (aa)-long G proteins. Alignments revealed more homogeneity within the European and North-American groups (at least 98 and 79% aa identity, respectively) than between European and North-American viruses (at best 70% a identity), and confirmed the presence of an extracellular divergent domain (positions 302-484) in APV-C G. A phylogenetic analysis demonstrated that North-American and French isolates of APV-C belonged to significantly different genetic lineages, in agreement with the different geographical origin and host species of these viruses. 相似文献
18.
Sequence of a bovine herpesvirus type-1 glycoprotein gene that is homologous to the herpes simplex gene for the glycoprotein gB 总被引:15,自引:0,他引:15
The 130-kDa bovine herpesvirus-1 (BHV-1) glycoprotein GVP 6 was found to cross-react immunologically with the herpes simplex glycoprotein gB. Antibodies in polyclonal serum against gB immunoprecipitated GVP 6 and its cleavage products from a lysate of BHV-1-infected cells. Conversely, polyclonal serum against GVP 6 precipitated gB from HSV-1-infected cell lysates. Sera against the other glycoproteins did not demonstrate cross-reactivity. A 3.6-kb Kpnl-Hpal fragment of BHV-1 DNA that hybridized to the gene for gB was cloned and the nucleotide sequence of both strands was determined. The longest codon reading frame in the fragment coded for a protein that showed extensive homology with gB1 and related sequences from pseudorabies virus, varicella-zoster virus, cytomegalovirus, and Epstein-Barr virus. The strongest homologies were observed in two segments of the ectodomain, the transmembrane domain, and sequences adjacent to the transmembrane domain. 相似文献