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为研究我国不同地区不同人群中HDV毒株的感染分子特征,从我国河南、内蒙、北京、四川、广西、西藏、新疆、辽宁、上海等地的HDV健康携带者、慢性丁肝病人与重症肝炎病人中筛选获得10余份HDV-RNA阳性血清。经逆转录一多聚酶链反应(RT-PCR)交叉扩增获得HDV抗原编码区的cDNA片段并克隆到PGEM-3Zf(-)或PGEM-T载体上,经序列分析研究其基因结构特点,结果表明:中国的HDV毒株基因型均为Ⅰ型,但至少存在ⅠA、ⅠB两个亚型,HDV毒株在不同地区间存在异质性,其中河南-1、-2、-3株及新疆株与台湾株同源性较高(核苷酸与氨基酸同源性分别大于92.1%与86.9%).当为ⅠA型;内蒙-1、四川、广西、西藏-1、辽宁、北京株与美国-1株同源性较高(核苷酸与氨酸同源性分别大于94.3%与88.8%),当为ⅠB亚型;上海株与意大利株的核昔酸同源性最高,为98.1%。研究证明我国新疆、内蒙、西藏等地区抗HD阳性率比其他省市高并不是由于存在其他基因型所致。 相似文献
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以采自河南的真菌传小麦花叶病毒(FWMV-C)为材料,抽提病毒RNA,合成互补DNA(cDNA)。对杂交筛选所得cDNA克隆进行亚克隆及序列分析,结果表明,亚克隆pGSI含有一个长度为891个核苷酸的不完整开放阅读框架(ORF)和长度为258个核苷酸的3'末端非编码区(NTR),并带有Poly(A)尾序。此段序列与大麦黄花叶病毒(BaYMV)及法国报道的一种小麦梭条花叶病毒(WSSMV-F)的RNA-13'末端分别具有67.6%和69.9%的同源性。由所测序列编码区(1-891nt)可推导产生296个氢基酸,并与WSSMV-F及BaYMV外壳蛋白氨基酸序列分别具有75.9%和71%的同源性。此结果表明,FWMV-C为另外一种不同于WSSMV-F的大麦黄花叶病毒组(Baymovirus)病毒,所测基因组片段应为RNA-13'末端序列,其中可能包括了病毒全长外壳蛋白编码区域。 相似文献
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北京等四地区呼吸道合胞病毒分离株G蛋白基因的比较分析 总被引:7,自引:0,他引:7
为了解我国不同地区呼吸道合胞病毒(RSV)感染特征是否与基因变异有关,对北京、广州、长春和河北四个地区不同流行特征中分离的RSV毒株的G蛋白基因进行了序列分析。结果表明,该G蛋白基因同国外A型亚原型株(A2株)间存在显著差异,A2株同分离株间的核苷酸同源性为92.7-93.6%,氨基酸同源性只有88.3-89.9%。分离株间的核苷酸同湃性为96.0-98.9%,氨基酸同源性92.6-97.7%。氨 相似文献
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黑子南瓜甘油-3-磷酸酰基转移酶基因的克隆及序列分析 总被引:6,自引:3,他引:3
依据国外报道的南瓜甘油-3-磷酸转酰酶(GPAT)基因的cDNA序列合成相应引物,用RT-PCR技术,成功地分离了黑子南瓜(Cucurbitaficifolia)GPAT基因的cDNA片段,并亚克隆到了pGEM-T载体系统的多克隆位点上,序列分析表明黑子南瓜GPAT基因的cDNA序列及递推的氨基酸序列与南瓜(Cucurbitamoschata)相比分别具有98%和965%的同源性。在1188bp中有22个核苷酸发生变化,导致13个氨基酸的改变 相似文献
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中国河南株丁型肝炎病毒全基因组的cDNA克隆和序列分析 总被引:3,自引:0,他引:3
从我国河南-抗丁型肝炎病毒抗原(anti-HDAg)及丁型肝炎病毒(HDv)RNA双阳性的HBsAg携带者血清中提取RNA,采用人工合成的引物进行逆转录和聚合酶链反应(PCR),获得了贯穿HDV全基因组的6个相互重叠的cDNA片段。经双脱氧末端终止法进行核苷酸序列分析,得到了长度为1674bp的我国人河南株HDVcDNA全序列。计算机分析表明,该株与我国台湾株(HDVIA型)、美国-1株(HDVIB型)、日本-1株(HDVⅡ型)和秘鲁-1株(HDVⅢ型)的核苷酸同源性分别为的94.3%、86.8%、75.4%和66.3%,氨基酸序列的同源性分别为89.7%、85.1%、71.9%和64.6%,并在核苷酸和推导的HDAg氨基酸序列中分别发现了5个和2个集中保守的区域。这些区域均与HDV的某些重要功能密切相关。 相似文献
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呼吸道合胞病毒在北京地区分离株G蛋白的基因分析 总被引:5,自引:0,他引:5
从经单克隆抗体证实为A亚型的北京地区呼吸道合胞病毒(RSV)分离株B79中,用RT-PCRT扩增出编码G蛋白的基因片段,克隆至载体pTZ18R中,经核苷酸序列测定证明,我国北京地区分离的A亚型株B79与RSVA亚型原型株(A2株)G蛋白基因的核苷酸同源性为93.8%,核苷酸的有义突变率为65%,由核苷酸推导出氨基酸序列的同源性为89.6%,氨基酸的变异主要集中在胞外区一个高度保守区的两端,而胞内区 相似文献
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用抗原捕获/多聚酶链反应(AC/PCR)对戊型肝炎病毒(HEV)细胞分离株MJ90和R25基因组的部分核苷酸序列进行扩增,获得了与HEV缅甸株ET1·1相同的cDNA扩增带。该cDNA扩增带纯化后用双脱氧核苷酸DNA链末端终止法测序,CJ90、R25株的核苷酸和氨基酸序列与ET1·1克隆的同源性分别为99.6%、100%和99%、99%,从而证明MJ90和R25毒株为HEV. 相似文献
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天花粉蛋白基因的克隆及序列分析 总被引:7,自引:0,他引:7
本文应用DNA多聚酶链式反应(PCR)技术,从括楼基因组DNA中扩增并克隆了天花粉蛋白(TCS)基因。核酸序列分析结果表明,克隆片段包括TCS的前原蛋白的编码序列和5'一侧翼区段。其编码序列与已发表的不同来源的3种TCS基因的核苷酸序列的同源性分别为99.20%,98.74%和98.64%。推导出的氨基酸序列与已发表的4种TCS的氨基酸序列的同源性分别为98.62%、98.62%、97.41%和9 相似文献
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鸡传染性支气管炎病毒免疫原基因cDNA的克隆和序列分析 总被引:3,自引:0,他引:3
从含有鸡传染性支气管炎病毒(IBV)的鸡胚尿囊液中快速提取病毒RNA,得到约23kb全长IBV RNA。运用反转录-聚合酶链反应(RT-PCR)得到1.72kb编码全长IBV免疫原糖蛋白S1的基因片段。进一步将此基因片段插入到质粒pUC19中,进行全长片段的序列分析。结果表明,IBV北京株和Beaudette株的核酸同源性为97.8%,与M41株的核酸同源性达98.5%。 相似文献
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采用RT-PCR方法合成了本研究室保存的番木瓜畸叶病毒(PMaLV)的外壳蛋白(CP)基因,将其CP基因克隆进Promega公司的pGEM-T and pGEM-T Easy Vector System(简称T-载体),并进行了序列分析。结果表明,PMaLV CP基因核苷酸序列全长为861nt,推导其编码287个氨基酸。与番木瓜环斑病毒(PRSV)美国夏威HA株系和澳大利亚W株系的CP基因相比,在第66nt处开始连续缺失3个核苷酸。与PRSV的华南Ys、Sm和G株系以及夏威夷的HA和澳大利亚的W株系相比,其CP基因序列同源率分别为96%、98%、95%、89%和89%。其的氨基酸序列同源率分别为98%、97%、97%、96%和95%。此结果表明,PMaLV属于PRSV的一个株系,不是一种新病毒。因此,我们称其为番木瓜环斑病毒畸叶株系(ML株系)。 相似文献
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Papaya ringspot potyvirus (PRSV) causes major diseases of papaya and cucurbits in the Indian subcontinent. Based on biological properties, PRSV isolates are classified as either papaya infecting (P), or non-papaya infecting (W) types. To characterise the P and W isolates from India at the molecular level, c. 1.7 Kb of the 3′-terminal regions comprising a part of the nuclear inclusion b (Nib) gene, the complete capsid protein (CP) gene and the untranslated region (UTR) of both the P and W isolates were cloned and sequenced. Comparative sequence analyses showed that the 3′-UTRs in isolates P and W were 209 nucleotides in length excluding the poly (A) tail, and shared 96% identity. The CP genes of the two isolates were also similar, with 87% nucleotide identity and 93% amino acid identity. The amino acid differences between the CP genes were mostly confined to the amino terminus. The DAG triplet associated with aphid transmissibility was present in the CP of isolate W, but it was replaced by DAD in the P isolate. The partially sequenced Nib genes were also 90% identical, but isolate W contained an additional amino acid (threonine) just upstream of the cleavage site (Q/S) between Nib and CP. This is the first reported comparison of the molecular characterisation of PRSV-P and W isolates from the Indian subcontinent. 相似文献
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Y. Sivaprasad A. Sujitha D.V.R. Sai Gopal 《Archives Of Phytopathology And Plant Protection》2013,46(2):172-179
Tobacco streak virus (TSV), a member of the genus Ilarvirus, family Bromoviridae is an important viral pathogen in peanut and other crops in South India. Fifteen TSV isolates naturally infecting groundnut, sunflower, onion, black gram, green gram, jute, tagetes, calotropis, pumpkin, watermelon and kenaf plants were collected from fields in different regions of Andhra Pradesh, Tamil Nadu and Karnataka. Virus was identified as TSV by direct antigen coating enzyme linked immunosorbent assay using TSV antiserum. The CP gene from each isolate was amplified using TSV coat protein specific primers. About 700 bp product was amplified, cloned, sequenced and determined its length as 717 nucleotides and codes for 239 amino acids. The sequence analysis revealed that the CP gene shared 91–100% and 91–99% sequence identity with TSV at nucleotide and amino acid level, respectively. The phylogenetic relationship based on the nucleotide sequence of these isolates from different geographical regions was also analysed in this study. 相似文献
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通过间接酶联免疫法(ID-ELISA)检测到染病落葵病样中存在黄瓜花叶病毒(Cucumber Mosaic Virus,CMV)。从病叶中提取总RNA,用RT-PCR方法扩增得到657bp的CMV CP基因片断,将扩增产物与T载体连接并进行测序。用DNA MAN将得到的CP基因序列与GenBank收录的黄瓜花叶病毒两亚组部分株系或分离物的CP基因序列进行比较,结果表明该CP基因与CMV亚组Ⅰ、亚组Ⅱ之间的核苷酸序列同源性分别为91.17~95.43%和75.30~75.76%,推导氨基酸序列同源性分别为95.41~97.71%和81.28~81.74%,表明CMV-Ba与亚组Ⅰ同源关系密切。 相似文献
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Natural infection of tospoviruses on three cucurbitaceous (Cucumis sativus, cucumber; Luffa acutangula, ridge gourd; Citrullus lanatus, watermelon) and three fabaceous (Vigna unguiculata, cowpea; Phaseolus vulgaris, French bean; Dolichos lablab, sem) vegetable crops in India was identified on the basis of nucleocapsid protein (NP) gene characteristics. The complete NP gene of the cowpea isolate from Kerala and the sem isolate from Tamil Nadu was 831 nucleotides long, encoding a protein of 276 amino acids. For other Tospovirus isolates from cucumber, French bean, ridge gourd and watermelon, the partial NP gene (291 nt) was sequenced. Comparative NP gene sequence analyses revealed that fabaceous isolates shared maximum identity both at the nucleotide (92–97%) and amino acid (93–97%) levels with the corresponding region of Groundnut bud necrosis virus (GBNV), whereas cucurbitaceous isolates shared maximum identity both at nucleotide (93–99%) and amino acid (95–98%) levels with the corresponding region of Watermelon bud necrosis virus (WBNV), results suggesting that the Tospovirus isolates infecting fabaceous hosts should be regarded as strain of GBNV, whereas those infecting cucurbitaceous hosts as a strain of WBNV. Nucleocapsid protein gene was conserved both in GBNV and WBNV isolates originating from different hosts and locations. 相似文献
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B.V. Bhaskara Reddy S. Obaiah L. Prasanthi Y. Sivaprasad A. Sujitha T. Giridhara Krishna 《Archives Of Phytopathology And Plant Protection》2013,46(4):345-346
To study the variability and to identify the species of Begomovirus associated with yellow mosaic disease of blackgram in Andhra Pradesh, India, infected blackgram samples were collected from six districts belonging to three regions of Andhra Pradesh. The total DNA was isolated by modified CTAB method and amplified with coat protein gene-specific primers (RHA-F and AC abut) resulting in 900?bp gene product. The PCR products were cloned, sequenced and deposited in GenBank. The sequence analysis of six clones showed that the size of amplified CP gene of YMV was 920?bp. Based on nucleotide sequence identity of six isolates representing three regions of Andhra Pradesh, the isolates from Rayalaseema and Telangana region are the same variant of YMV (>99.5% identity) and isolate from coastal Andhra is another variant of YMV (>95.4%) when compared with other region isolates. Comparison of CP gene sequence of YMV-TPT isolate with 27 other isolates in database revealed more than 93.2 and 86.2% identity with MYMIV isolates and less than 80 and 64% identity with MYMY isolates that originate from Indian sub-continent and South-East Asia at nucleotide and amino acid level, respectively. Phylogenetic tree based on CP gene sequences of six isolates with other isolates from GenBank formed unique cluster with MYMIV. Hence the YMV infecting blackgram in Andhra Pradesh is caused by MYMIV rather than MYMY as reported in Tamil Nadu which is adjoining state in southern India. 相似文献
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CHENG Zhuomin HE Xiaoyuan WU MaosenZHOU Guanghe(Paul Keese P. M. Waterhouse 《中国科学:生命科学英文版》1996,39(5):534-543
GPV is a Chinese serotype isolate of barley yellow dwarf virus (BYDV) that has no reactionwith antiserum of MAV, PAV, SGV, RPV and RMV. The sequence of the coat protein (CP) of GPV isolate of BYDV was identified and its amino acid sequence was deduced. The coding region for the putative GPV CP is 603 bases nucleotides and encodes a Mr 22218 (22 ku) protein. The same as MAV, PAV and RPV, GPV contained a second ORF within the coat protein coding region. This protein of 17024 Mr (17 ku) is thought to correspond to the Virion protein genome linked (Vpg). Sequence comparisons of the CP coding region between the GPV isolate of BYDV and other isolates of BYDV have been done. The nucleotide and ammo acid sequence homology of GPV has a greater identity to the sequence of RPV than those of PAV and MAV. The GPV CP sequence shared 83.7% of nucleotide similarity and 77.5% of deduced amino add similarity, whereas that of the PAV and MAV shared 56.9%. 53.2% and 44.1%. 43.8% respectively. According to BYDV-GPV CP seque 相似文献
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家蚕浓核病毒(镇江)株主要结构蛋白基因的克隆及表达 总被引:3,自引:0,他引:3
家蚕浓核病毒(Bombyx mori densovirus,BmDNV)是一种昆虫细小病毒.与其它昆虫细小病毒感染昆虫体内多种组织不同,家蚕浓核病毒只感染家蚕中肠上皮组织的圆筒型细胞,感染该病毒细胞的细胞核可以被孚尔根和甲基绿浓染,在病毒感染的早期中肠上皮组织细胞数量增加,形成褶皱,最后感染细胞脱落到肠腔中[1-3].自从20世纪70年代末日本学者证实家蚕浓核病是由于家蚕浓核病毒感染引起的以来[4],已经分离得到了多个病毒株系[5-8].根据它们在血清学、理化特性、品种感受性和病理特征等方面的差异,分为BmDNV-1(伊那株)和BmDNV-2(以山梨株为代表)[8-11]. 相似文献