共查询到18条相似文献，搜索用时 187 毫秒
1.
用鸡源新城疫病毒凤阳分离株WF00C对9～10 日龄SPF鸡胚的尿囊腔接种,成功增殖了该病毒,采用一步法RT-PCR技术扩增WF00C病毒的F基因,获得了1条1.7kb的特异性条带.用PCR产物直接测序.测序结果表明,扩增片段大小为1782bp,含有1个1662bp的开放性阅读框,编码554个氨基酸.核苷酸同源性分析表明:WF00C株与国内外其他NDV F基因的同源性为84.8%～97.5%,其中与国内标准强毒株F48E9的同源性为87.1%,说明WF00C与国内外的传统毒株有较大变异.与Taiwan95株和J株的同源性为94.1%和97.5%,说明WF00C与Taiwan95株和J株亲缘关系较近,具有较高的相似性.F蛋白裂解位点的氨基酸顺序为112Arg-Arg-Gln-Lys -Arg-Phe117, 表明为NDV的强毒株.蛋白疏水性和抗原性分析表明与标准强毒株相比没有太大的变异. 相似文献
2.
用鸭源WF01D株新城疫病毒接种对9-10日龄SPF鸡胚尿囊腔,成功增殖了该病毒,采用一步法RT-PCR技术扩增WF01D病毒的HN基因,获得了1条约1.8kb的特异性条带。PCR产物回收纯化后测序。测序结果表明,扩增片段大小为1844bp,含有1个1716bp的开放性阅读框,编码571个氨基酸。核苷酸同源性分析表明：WF01D与国内外其他NDV HN基因的同源性为81.2%-95.0%,其中与国内标准强毒株F48E9的同源性为84.3%,说明WF01D与国内外的传统毒株有较大变异。与Taiwan95株和NL/96株的同源性为93.8%和95.0%,说明WF01D与Taiwan95株和NL/96株亲缘关系较近,具有较高的相似性。 相似文献
3.
用RT？PCR方法从云南省寻甸、嵩明、通海、芒市、沾益和陆良6个地区的水稻样品中扩增到水稻矮缩病毒S10片段部分序列,核苷酸测定及序列分析表明,6个地区水稻矮缩病毒分离物S10片段部分序列长度均为1 071个核苷酸,含1个1 059个核苷酸的ORF,编码352个氨基酸.云南分离物S10片段之间的核苷酸同源性约为97.9%~99.9%,其编码的氨基酸同源性为97.7%~100%;云南分离物与日本分离物的S10片段核苷酸同源性和氨基酸相似性分别为94.1%~95.4%和96.3%~99.2%,与中国分离物的S10片段核苷酸同源性和氨基酸相似性分别为94.8%~96.6%和95.2%~96.6%.云南分离物在亲缘关系上与日本分离物较中国分离物近. 相似文献
4.
根据Imbereehts等报道的EHEC F18菌毛主要亚单位(fedA)基因序列设计一对引物，以重庆地区仔猪水肿病流行强毒株W96基因组DNA为模板，采用聚合酶链式反应(PCR)技术扩增到约500bp的核酸片段。用常规DNA重组技术将该片段克隆到表达载体pET28b( )的SalI-Hindm位点之间构建重组表达质粒pET28fedA，序列测定结果显示该克隆片段长447bp，同源性分析表明该基因片段与GenBank报道的fedA核苷酸序列有99．4％的同源性，W96株fedA基因三处核苷酸发生变异，其中两个为无义突变，另一个导致Q100R突变。将pET28fedA转入BL21(DE3)中进行诱导表达，经SDS—PAGE电泳检测上清存在约20KD的蛋白，6xHis affinity tag IMAC亲和层析显示该蛋白能被亲和纯化，说明是含有组氨酸标签的融合目的蛋白。 相似文献
5.
茶树β-1,3-葡聚糖酶基因cDNA片段的克隆与序列分析 总被引：2，自引：0，他引：2
文章首次从茶树中克隆出-β1,3-葡聚糖酶基因cDNA1369bp连续序列。根据已发表的植物中β-1,3-葡聚糖酶基因的保守序列,设计一对简并引物,采用RT-PCR技术,从茶树中扩增出366bp的cDNA片段。根据此片段序列设计特异引物,利用3'RACE获得-β1,3-葡聚糖酶cDNA 3'端1252bp的cDNA片段。序列分析表明:该基因cDNA 3'端核苷酸序列及其推测的氨基酸序列与其他植物的-β1,3-葡聚糖酶基因家族的cDNA相应序列同源性为50%-90%,经序列拼接,本研究从茶树中克隆出-β1,3-葡聚糖酶基因cDNA3'端1369bp的连续序列,GenBank登录号为AF399920。 相似文献
6.
CMV甜瓜分离物外壳蛋白基因的克隆及其生物信息学分析 总被引：1，自引：0，他引：1
马新艳 《南阳师范学院学报》2008,7(12)
从河南省临颖县采集的病毒感染的甜瓜样本经ELISA检测和接种分离获得黄瓜花叶病毒（Cucumber mosaic vi-rus,CMV）分离物。把该分离物接种于西葫芦,从发病的叶片中提取总RNA,并以此为模板经RT-PCR扩增获得CMV的外壳蛋白（cp）基因,将其克隆到pUCm-T质粒上。经序列测定和生物信息学分析,结果表明该cp基因由657个核苷酸组成,编码218个氨基酸。预测该蛋白的等电点为5。12,分子量约为24kD。其核苷酸序列与黄瓜花叶病毒亚组I的分离物有较高的同源性,达92.2%～93.9%,与亚组II的同源性仅为76.8%～77.8%,与我国报道的CMV分离物的cp基因序列比较,只有香蕉株系XB外核苷酸序列的同源性达91.8%～93.4%。根据这些分析,该CMV分离物属于亚组I。另外还对黄瓜花叶病毒外壳蛋白的高级结构作出了预测。 相似文献
7.
通过对犬冠状病毒TN449株纤突蛋白基因的克隆测序,将获得的序列与GeneBank中登陆的犬冠状病毒不同毒株S基因序列进行比较,分析它们的同源性、差异程度,同时绘制核苷酸序列及其氨基酸序列的系统进化树.有助于了解犬冠状病毒的抗原变异、血清型和毒力变化规律. 相似文献
8.
黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus, CGMMV)是葫芦科重要种传病毒之一.本文建立了免疫捕获RT-PCR(IC-RT-PCR)的分子检测方法直接从带毒种子中检测出该病毒,扩增获得其CP基因,并克隆到pMD18-T载体中,经核苷酸序列分析表明,该分离物CP基因全长为486个核苷酸,编码由161个氨基酸组成的17.3kDa蛋白,与国内外已报道的CGMMV的CP基因相比,其核苷酸序列的同源性为91.4%~99.4%,其推导的氨基酸同源性为98.8%~100%.IC-RT-PCR方法的建立,为检疫部门提供了从带毒种子中快速、准确检测该病毒的方法,很有应用前景. 相似文献
9.
采用高保真PCR从我国钩体黄疸出血群赖株基因组中扩增全长mviN基因片段,对钩体黄疸出血群赖株mviN基因进行克隆.结果显示,所克隆的mviN基因的核苷酸和氨基酸序列与已发表的mviN基因序列(Accesion No.in GenBank: NC004343)同源性分别为99.68%和99.42%.因此可以认为,我们成功克隆了mviN基因,并为该基因的后续研究奠定了基础. 相似文献
10.
11.
Ling-zhe Huang Li-xia Rao Xue-ping Zhou Jian-xiang Wu 《Journal of Zhejiang University. Science. B》2013,14(10):875-885
Rice stripe virus (RSV) is the type member of the genus Tenuivirus. RSV is known to have four segmented, single-stranded RNA molecules and causes rice stripe disease in the rice fields of China, Japan, and Korea. Based on the complete genomic sequences of the determined 6 RSV isolates (from Yunnan, Jiangsu, Zhejiang, and Liaoning Provinces, China) and 27 other RSV isolates (from Yunnan, Jiangsu, Anhui, Henan, and Shandong Provinces of China, also Japan and Korea) downloaded from GenBank, we provided a genotyping profile of RSV field isolates and described the population structure of RSV. All RSV isolates, except isolate CX, could be divided into two subtypes, one including 6 isolates from Yunnan Province, and the other including 26 isolates from different parts of China, Japan, and Korea, which were referred to as subtype II and subtype I, respectively. The amino acid distances between subtypes range from 0.053 to 0.085. RSV isolates in Yunnan Province were genetically differentiated from other parts of China, Japan, and Korea and showed infrequent gene flow. The RSV populations collected from other parts of China, Japan, and Korea were only composed of subtype I and showed very low genetic diversity. We speculated that isolate CX may be the result of recombination of isolates from two subtypes. Two potential recombination events were detected in RNA4 of isolate CX. 相似文献
12.
Broad bean wilt virus 2 (BBWV2) isolate P158 was characterized and the complete nucleotide sequence of viral genomic RNA2
was determined. P158 has 30 nm diameter isometric particles. Double immunodiffusion test suggested that P158 has high antigenicity
homology with BBWV2 isolate B935. SDS-PAGE result showed that the coat protein of P158 was comprised of two types of polypeptide
with molecular weight of 44.7 kD and 21.9 kD, respectively. The genome of P158 was made up of two RNA molecules with the length
of 6.0 kb and 3.6 kb, respectively. cDNA of RNA2 was cloned by RT-PCR and sequenced. RNA2 was composed of 3597 nucleotide
(nt) residues excluding the poly(A) tail and contained single long open reading frame extending from nt 230 to nt 3424 in
the viral sense RNA, and encoded a polyprotein ofMr 119002 (119K). Comparison of the polyprotein and the counterpart of isolate B935 indicated that the polyprotein was cleaved
at Q/G (465/466) and Q/A (867/868), to release three mature proteins: a protein of unknown function, large and small subunit.
Sequence comparisons of P158 with fabaviruses showed P158 had very high sequence homology with BBWV2 isolates and patchouli
mild mosaic virus, but to a less extent with BBWV1 isolates.
Project supported by NSFC (No 39770035; 39900005). The GenBank accession number of the sequence reported in this paper is
AF228423. 相似文献
13.
根据GenBank发表的鸡γ-干扰素核苷酸序列,使用primer 5设计一对特异性引物,通过RT-PCR技术从ConA诱导培养的鸡脾脏淋巴细胞中克隆出鸡γ-干扰素基因并对其进行测序,测序结果表明,鸡γ-干扰素基因全长495bp,具有一个完整的开放阅读框,编码164个氨基酸,与国外发表的序列比较,两序列间同源性为100%.计算机软件对鸡γ-干扰素编码的氨基酸序列进行了抗原性分析,结果表明鸡γ-干扰素具有良好的免疫原性. 相似文献
14.
ＩＮＴＲＯＤＵＣＴＩＯＮＢｒｏａｄｂｅａｎｗｉｌｔｖｉｒｕｓ (ＢＢＷＶ)ｉｓｔｈｅｔｙｐｅｍｅｍｂｅｒｏｆｔｈｅｇｅｎｕｓＦａｂａｖｉｒｕｓ.Ｉｔｈａｓａｗｉｄｅｈｏｓｔｒａｎｇｅａｍｏｎｇｄｉｃｏｔｙｌｅｄｏｎｓａｎｄｓｏｍｅｆａｍｉｌｉｅｓｏｆｍｏｎｏｃｏｔｙｌｅｄｏｎｓ,ａｎｄｉｓａｎｅｃｏｎｏｍｉｃａｌｌｙｉｍ ｐｏｒｔａｎｔｖｉｒｕｓｉｎＣｈｉｎａ (Ｚｈｏｕｅｔａｌ.,1 994 ) .ＢＢＷＶｈａｓｉｓｏｍｅｔｒｉｃｐａｒｔｉｃｌｅｓ,ｈｅｘａｇｏｎａｌｉｎｏｕｔ ｌｉｎｅａｎｄ 3 0ｎｍｉｎｄｉａｍ… 相似文献
15.
Robinson David Jebakumar SOLOMON Amit KUMAR Velayudhan SATHEEJA SANTHI 《Journal of Zhejiang University. Science. B》2013,14(12):1162-1172
Atrazine is a selective herbicide used in agricultural fields to control the emergence of broadleaf and grassy weeds. The persistence of this herbicide is influenced by the metabolic action of habituated native microorganisms. This study provides information on the occurrence of atrazine mineralizing bacterial strains with faster metabolizing ability. The enrichment cultures were tested for the biodegradation of atrazine by high-performance liquid chromatography(HPLC) and mass spectrometry. Nine cultures JS01.Deg01 to JS09.Deg01 were identified as the degrader of atrazine in the enrichment culture. The three isolates JS04.Deg01, JS07.Deg01, and JS08.Deg01 were identified as efficient atrazine metabolizers. Isolates JS04.Deg01 and JS07.Deg01 produced hydroxyatrazine(HA) N-isopropylammelide and cyanuric acid by dealkylation reaction. The isolate JS08.Deg01 generated deethylatrazine(DEA), deisopropylatrazine(DIA), and cyanuric acid by N-dealkylation in the upper degradation pathway and later it incorporated cyanuric acid in their biomass by the lower degradation pathway. The optimum pH for degrading atrazine by JS08.Deg01 was 7.0 and 16S rDNA phylogenetic typing identified it as Enterobacter cloacae strain JS08.Deg01. The highest atrazine mineralization was observed in case of isolate JS08.Deg01, where an ample amount of trzD mRNA was quantified at 72 h of incubation with atrazine. Atrazine bioremediating isolate E. cloacae strain JS08.Deg01 could be the better environmental remediator of agricultural soils and the crop fields contaminated with atrazine could be the source of the efficient biodegrading microbial strains for the environmental cleanup process. 相似文献
16.
采用鸡胚接种法从安徽凤阳某患病鹅群中分离到一株病毒。经HA与HI试验、血清中和接种鸡胚试验、病毒回归试验确认为鹅副粘病毒 ,并命名为WF0 0 G株。参照新城疫病毒毒力判定的标准及其方法 ,测定该分离株的鸡胚最小致死量平均死亡时间 (MDT)、1日龄鸡脑接种致病指数 (ICPI)和 6周龄鸡静脉内接种致病指数 (IVPI)分别为 4 4 8h、1 81和 2 32 ,结果表明该分离株具有与新城疫病毒 (NDV)速发型相类似的毒力 ,属强毒力毒株。 相似文献
17.
Ya-e Zhao Zheng-hang Wang Yang Xu Ji-ru Xu Wen-yan Liu Meng Wei Chu-ying Wang 《Journal of Zhejiang University. Science. B》2012,13(10):763-768
To our knowledge, few reports on Demodex studied at the molecular level are available at present. In this study our group, for the first time, cloned, sequenced and analyzed the chitin synthase (CHS) gene fragments of Demodex folliculorum, Demodex brevis, and Demodex canis (three isolates from each species) from Xi’an China, by designing specific primers based on the only partial sequence of the CHS gene of D. canis from Japan, retrieved from GenBank. Results show that amplification was successful only in three D. canis isolates and one D. brevis isolate out of the nine Demodex isolates. The obtained fragments were sequenced to be 339 bp for D. canis and 338 bp for D. brevis. The CHS gene sequence similarities between the three Xi’an D. canis isolates and one Japanese D. canis isolate ranged from 99.7% to 100.0%, and those between four D. canis isolates and one D. brevis isolate were 99.1%–99.4%. Phylogenetic trees based on maximum parsimony (MP) and maximum likelihood (ML) methods shared the same clusters, according with the traditional classification. Two open reading frames (ORFs) were identified in each CHS gene sequenced, and their corresponding amino acid sequences were located at the catalytic domain. The relatively conserved sequences could be deduced to be a CHS class A gene, which is associated with chitin synthesis in the integument of Demodex mites. 相似文献
18.
目的：克隆蚓激酶基因并在GENEBANK中进行序列分析．方法：采用RT—PCR技术，以蚯蚓总．RNA为模板进行扩增，克隆蚓激酶基因、用Blast软件对基因进行同源性分析．结果：克隆了一个cDNA片段，与GENEBANK中蚓激酶基因序列最高同源性为99％．结论：本方法实用可行，同源性分析表明克隆的cDNA片段具备完整的蚓激酶编码区，为下一步进行蚓激酶的表达研究奠定了良好的基础． 相似文献