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1.
为了获得2009年新型甲型H1N1流感病毒与2008流感病毒的基因序列及氨基酸序列的一致性,以便对一个基因家族的生物学特征有一个简明扼要的了解,针对目前流行的新型甲型H1N1流感病毒的基因序列及其所编码的氨基酸序列,采用动态规划算法对其一级序列进行序列相似度分析,获得了2009年新型甲型H1N1流感病毒的NA和M基因片段以及2008年猪源性甲型H1N1流感病毒的相应基因片段同源性高、在有些位点发生了基因突变增添和突变缺失等重要基因信息。为此次新型甲型H1N1流感病毒的研究提供了依据。  相似文献   

2.
甲型H1N1流感病毒(influenzavirusA)基因组合有8个RNA基因片段,本文主要介绍了RNA的结构,以及目前计算机在RNA结构分析领域的应用现状,以及应用最多的预测工具。  相似文献   

3.
流行性感冒的分子流行病学研究进展   总被引:1,自引:0,他引:1  
杨红 《甘肃科技纵横》2003,32(5):127-127,118
流行性感冒实质上是一种人畜共患的传染病,至今仍无法有效控制其流行。人群流感大流行主要是由于甲型流感病毒抗原变异所致,且甲型流感病毒抗原的基因结构可能在人、禽、畜问发生重配和重组,引起人、禽、畜之问的传播并发生流行。通过积极有效地监测流感病毒在人和家禽、家畜中的流行特点,可以预测将要出现的流感病毒毒株,以制备相应的疫苗,达到控制流感流行的目的。  相似文献   

4.
H5N1亚型禽流感病毒M基因的克隆与分子进化分析   总被引:1,自引:0,他引:1  
以一株H5N1亚型禽流感病毒RNA为模板,用RT-PCR方法,扩增M基因全长,将PCR产物克隆于pMD18-T载体,测序结果表明所克隆的982个核苷酸的片段包含了M1和M2基因的完整阅读框架,通过软件推导M1和M2基因分别编码252和97个氨基酸,将M全长序列与Genbank收录的10株H5N1亚型流感病毒M基因序列进行比较,病毒株之间M基因核苷酸序列同源性为92.3%~99.3%,编码的两个蛋白M1和M2氨基酸序列间同源性分别为为96.0%~98.8%和92.9%~99%,分子进化分析揭示了病毒株间的亲源关系.  相似文献   

5.
本文报道了印鼠客蚤云南株线粒体DNA中长为901bp的片段,包括完整的COⅡ基因和3个氨基酸tRNA基因及ATPase8基因片段。COⅡ基因全长684bp,编码227个氨基酸,起始密码为ATC,终止密码为TAA。印鼠客蚤mtDNACOⅡ基因中富含AT,含量为77%,GC含量为23%。根据COⅡ基因核苷酸和氨基酸序列,对蚤目蚤科部分种类及外群进行了分子系统学分析。  相似文献   

6.
一株鸡副粘病毒的分子特性研究   总被引:28,自引:1,他引:27  
应用RT-PCR技术对新城疫病毒(NDV) Y98株的F基因重要功能区片段扩增后进行序列测定,并与多株已报导的NDV参考株相应片段进行序列比较,经遗传基因进化树分析后,首次报道了基因Ⅶ型NDV分离株在中国大陆的存在.  相似文献   

7.
采用RT-PCR方法获得了青岛文昌鱼2497bp长的hedgehog基因片段,其所编码的氨基酸序列与多种生物hh基因家族成员的相应片段显示了较高的同源性.同时获得2个含有部分hedgehog基因片段的序列hedgehog-like-1和hedgehog-like-2,提示文昌鱼中发生hedgehog基因倍增过程和有多拷贝hh基因存在的可能性.为研究hh基因的分子进化提供了线索.  相似文献   

8.
采用噬菌体生物扩增法对本实验室分离鉴定的25株奶牛结核分支杆菌进行药敏分析,检测出14株耐链霉素、利福平、异烟肼菌株.用PCR扩增耐药株的rpsL基因片段(370bp)、rpoB基因片段(213bp)和KatG基因片段(458bp),并对目的片段进行测序分析,其中5株耐链霉素菌株的rpsL基因在43位点发生突变,均由赖氨酸密码子(AAG)突变为精氨酸密码子(AGG);5株利福平耐药株的rpoB基因在531位点、526位点、5¨位点发生突变,其中有3株分离株的rpoB基因在531位点发生点突变,1株分离株的rpoB基因在526位点发生突变,1株分离株的rpoB基因在531位点和511位点发生了较为少见的联合突变;4株异烟肼耐药菌株,其中2株在315位点发生碱基突变,2株在463位点发生碱基突变.  相似文献   

9.
甲型H1N1流感病毒基因测序完成 加拿大科学家已经完成对3个甲型H1N1流感病毒样本的基因测序,这是世界上首次完成对这种新病毒的基因测序,将为研制疫苗打下基础。  相似文献   

10.
陈小玲  陈东  芦志龙 《广西科学》2011,18(3):264-268
为了提高瑞氏木霉(Trichoderma reesei)的纤维素酶活力,用分子改造的方法改造其内切-β-1,4葡聚糖酶基因Egl 1.用DNase I消化EglL 1,回收50~200bp的片段,用T4 DNA连接酶连接回收的片段,进行PCR反应,将PCR产物转入瑞氏木霉原生质体,用比较滤纸酶活力的方法筛选纤维素酶活力...  相似文献   

11.
Subtypes of H1N1 influenza virus can be found in humans in North America, while they are also associated with the infection of swine. Characterization of the genotypes of viral strains in human populations is important to understand the source and distribution of viral strains. Genomic and protein sequences of 10 isolates of the 2009 outbreak of influenza A (H1N1) virus in North America were obtained from GenBank database. To characterize the genotypes of these viruses, phylogenetic trees of genes PB2, PB1, PA, HA, NP, NA, NS and M were constructed by Phylip3.67 program and N-Linked glycosylation sites of HA, NA, PB2, NS1 and M2 proteins were analyzed online by NetNGlyc1.0 program. Phylogenetic analysis indicated that these isolates are virtually identical but may be recombinant viruses because their genomic fragments come from different viruses. The isolates also contain a characteristic lowly pathogenic amino acid motif at their HA cleavage sites (IPSIQSR↓GL), and an E residue at position 627 of the PB2 protein which shows its high affinity to humans. The homologous model of M proteins showed that the viruses had obtained the ability of anti-amantadine due to the mutation at the drug-sensitive site, while sequence analysis of NA proteins indicated that the viruses are still susceptible to the neuraminidase inhibitor drug (i.e. oseltamivir and zanamivir) because no mutations have been observed. Our results strongly suggested that the viruses responsible for the 2009 outbreaks of influenza A (H1N1) virus have the ability to cross species barriers to infect human and mammalian animals based on molecular analysis. These findings may further facilitate the therapy and prevention of possible transmission from North America to other countries.  相似文献   

12.
目的克隆、表达和鉴定流感病毒H3N2血凝素基因(hemagglutinin,HA)和神经氨酸酶基因(neuramidinase,NA)序列,为制备抗体和基因工程疫苗打下基础。方法在成功克隆流感病毒H3N2全长HA、NA基因并测序的基础上,将部分基因序列克隆到表达载体pMET A上,构建了重组表达质粒pMET A/HA(52 bp~1 549 bp)、pMET A/NA(121 bp~1 260 bp),电转化真核酵母菌pMAD16,甲醇诱导表达,利用Ni2+亲和层析柱对重组蛋白进行纯化,并用Western Blotting和ELISA方法检测其抗原性。结果重组蛋白在酵母菌中可以高效表达,SDS-PAGE显示蛋白表达后形成了二聚体,蛋白纯度占总蛋白的95%以上,ELISA和Western Blotting实验证实,重组蛋白具有良好的抗原性。结论本研究成功克隆和表达了流感病毒H3N2 HA、NA基因序列,为流感病毒H3N2诊断试剂和疫苗开发等进一步研究奠定了基础。  相似文献   

13.
目的克隆、表达和鉴定禽流感病毒H5N1血凝素基因(hemagglutinin,HA)和神经氨酸酶基因(neuramidinase,NA)序列,为制备抗体和基因工程疫苗打下基础。方法在成功克隆禽流感病毒H5N1全长HA、NA基因并测序的基础上,将部分基因序列克隆到表达载体pMET A上,构建了重组表达质粒pMET A/HA(49~1 587 bp)、pMET A/NA(121~1 200 bp),电转化真核酵母菌pMAD16,甲醇诱导表达,利用Ni2+亲和层析柱对重组蛋白进行纯化,并用Western Blotting和ELISA方法检测其抗原性。结果重组蛋白在酵母菌中可以高效表达,SDS-PAGE显示蛋白表达后形成了二聚体,蛋白纯度占总蛋白的95%以上,ELISA和Western Blotting实验证实,重组蛋白具有良好的抗原性。结论本研究成功克隆和表达了禽流感病毒H5N1 HA、NA基因序列,为禽流感病毒H5N1诊断试剂和疫苗的开发等进一步的研究提供了依据。  相似文献   

14.
The molecular and pathogenic properties of avian influenza virus (A/duck/Hubei/216/1985/H7N8) isolated from Hubei Province of China in 1985 were characterized.The hemagglutinin gene (HA) of Dk/Hb/216/85/H7N8 had the multiple amino acid se-quences (-PEIPKGRG-) at the connecting peptide between HA1 and HA2, which is considered to be a distinguishing molecular characteristic of low pathogenicity.The key sites of host markers among the genes (M, NP, NS, PA and PB2) of Dk/Hb/ 216/85/H7N8 were similar to those of...  相似文献   

15.
Since the 2009 pandemic H1N1 swine-origin influenza A virus (09 S-OIV) has reminded the world about the global threat of the ever changing influenza virus,many questions regarding the detailed re-assortment of influenza viruses yet remain unanswered.Influenza A virus is the causative agent of the pandemic flu and contains 2 major antigenic glycoproteins on its surface:(i) hemagglutinin (HA);and (ii) neuraminidase (NA).The structures of the 09 S-OIV HA and NA proteins (09H1 and 09N1) have recently been resolved in our laboratory and provide some clues as to why the 09 S-OIV re-assortment virus is highly infectious with severe consequences in humans.For example,the 09H1 is highly similar to the HA of the 1918 influenza A pandemic virus in overall structure and especially in regards to its 5 defined antibody binding epitopes.For 09N1,its most distinctive feature is the lack of a 150-loop active site cavity,which was previously predicted to be present in all N1 NAs,and we hypothesize that the 150-loop may play a important role in the substrate specificity (α2,3 or α2,6 linked sialic acid receptors) and enzymatic mechanism of influenza NA.Combination of the HA and NA with special characteristics for the 09 S-OIV might contribute to its high increased transmissibility in humans.  相似文献   

16.
Here we report the codon bias and the mRNA secondary structural features of the hemagglutinin (HA) cleavage site basic amino acid regions of avian influenza virus H5N1 subtypes. We have developed a dynamic extended folding strategy to predict RNA secondary structure with RNAstructure 4.1 program in an iterative extension process. Statistical analysis of the sequences showed that the HA cleavage site basic amino acids favor the adenine-rich codons, and the corresponding mRNA fragments are mainly in the folding states of single-stranded loops. Our sequential and structural analyses showed that to prevent and control these highly pathogenic viruses, that is, to inhibit the gene expression of avian influenza virus H5N1 subtypes, we should consider the single-stranded loop regions of the HA cleavage site-coding sequences as the targets of RNA interference.  相似文献   

17.
He X  Zhou J  Bartlam M  Zhang R  Ma J  Lou Z  Li X  Li J  Joachimiak A  Zeng Z  Ge R  Rao Z  Liu Y 《Nature》2008,454(7208):1123-1126
  相似文献   

18.
The epidemic situation of A H1N1 flu arose in North America in April 2009, which rapidly expanded to three continents of Europe, Asia and Africa, with the risk ranking up to 5. Until May 13th, the flu virus of A H1N1 had spread into 33 countries and regions, with a laboratory confirmed case number of 5728, including 61 deaths. Based on IRV and EpiFluDB database, 425 parts of A H1N1 flu virus sequence were achieved, followed by sequenced comparison and evolution analysis. The results showed that the current predominant A H1N1 flu virus was a kind of triple reassortment A flu virus: (i) HA, NA, MP, NP and NS originated from swine influenza virus; PB2 and PA originated from bird influenza virus; PB1 originated from human influenza virus. (ii) The origin of swine influenza virus could be subdivided as follows: HA, NP and NS originated from classic swine influenza virus of H1N1 subtype; NA and MP originated from bird origin swine influenza virus of H1N1 subtype. (iii) A H1N1 flu virus experienced no significant mutation during the epidemic spread, accompanied with no reassortment of the virus genome. In the paper, the region of the representative strains for sequence analysis (A/California/04/2009 (H1N1) and A/Mexico/4486/2009 (H1N1)) included USA and Mexico and was relatively wide, which suggested that the analysis results were convincing.  相似文献   

19.
The neuraminidase (NA) in viral surface is one of the main subtype-specific antigen of influenza type A viruses.Neuraminidase is an enzyme to break the bonds between hemagglutinin (HA) and sialic acid to release newly formed viruses from infected cells.In this study,the H3N2 subtype virus NA genes were sequenced and NA proteins were screened for B-cell epitopes and assessed based on immunoinformatics.Based on this results,four peptides DR6,EY7,VG8 and RE8 (covering amino acid residues 151-156,368-374,398-405 and 428-435,respectively) of the NA protein were synthesized artificially.These peptides were used to immunize New Zealand rabbits subcutaneously to raise antisera.Experimental results showed that these four peptides were capable of eliciting antibodies against H3N2 viruses in a specific and sensitive feature,detected in vitro by enzyme-linked immunosorbent assay.Moreover,hemadsorption anti-releasing effects took place in three three-antisera-mixtures at a dilution of 1:40.Alignment using NA gene database showed that amino acid residues in these four epitope peptides were substituted at specific sites in all the NAs sequenced in this study.It was suggested that these NA epitope peptides might be used in combination with HA proteins as vaccine antigens.  相似文献   

20.
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