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1.
参照GenBank中内源性绵羊肺腺瘤病毒enJS56A1株pol基因序列设计了2对引物。应用PCR技术特异性地扩增出病毒的pol基因片段,将其克隆到PMD19-T载体后测序。应用计算机软件将测定序列与内源性南非代表毒株enJS56A1(AF153615)的pol基因序列比较,核苷酸同源性为99.3%,推导出的氨基酸同源性为95.0%。与外源性南非代表株JSRV-SA(M80216)的pol基因序列比较,核苷酸同源性为99.0%,氨基酸同源性为99.3%。这也是我国首次报道的内源性绵羊肺腺瘤病毒pol基因序列,为我国科研工作者进行更深入的研究奠定基础。 相似文献
2.
内源性绵羊肺腺瘤病毒NM株gag基因的克隆、序列分析及蛋白结构预测 总被引：2，自引：0，他引：2
提取内源性绵羊肺腺瘤病毒内蒙古分离株（NM）总DNA，参照GenBank中内源性绵羊肺腺瘤病毒enJS56A1株gag基因序列设计1对引物。应用PCR技术特异性地扩增出病毒的gag基因片段，将其克隆到pMD19-T载体中进行测序得到完整的gag基因序列，并用DNAStar软件进行序列分析，分析结果表明，与内源性南非代表毒株enJS56A1（AF153615）的gag基因序列比较，核苷酸同源性为98．9％，推导出的氨基酸同源性为98．4％。与外源性美国代表株JSRV21（AF105220）的gag基因序列比较，核苷酸同源性为89．6％，氨基酸同源性为94．8％。利用生物信息学软件对其蛋白结构进行预测，结果表明gag-enJSRV-NM为一结构松散的蛋白分子，这也是我国首次报道的内源性绵羊肺腺瘤病毒的gag基因的全序列，为我国科研工作者进行更深入的研究奠定了基础。 相似文献
3.
参照内源性绵羊肺腺瘤病毒株enJSRV-20(EF680302.1)全基因组序列设计合成3对引物,以山羊基因组DNA为模板,应用PCR技术扩增山羊内源性病毒基因片段,分别连接PMD-18T载体并测序,完成了1株山羊内源性肺腺瘤反转录病毒全基因序列的测定。结果分析显示测定序列全长7 480bp,获得的序列与内源性绵羊肺腺瘤病毒株enJSRV-18(EF680301.1)的核苷酸相似性为93.4%,与外源性绵羊肺腺瘤病毒株JS-7(AF357971.1)的核苷酸相似性为88.1%。根据pro基因序列构建系统进化树,结果显示扩增序列与内源性绵羊反转录病毒株en-JSRV-5(EF680305.1)的亲缘关系最近。本研究有助于内源性肺腺瘤反转录病毒感染性克隆的构建和内源性肺腺瘤反转录病毒功能的研究。 相似文献
4.
参照内源性绵羊肺腺瘤病毒株enJSRV-20全基因组序列设计合成3对引物,应用PCR技术扩增内源性绵羊肺腺瘤病毒基因片段分别连接pMD-18T载体并测序,完成了内源性绵羊反转录病毒基因组序列测定。分析结果显示,测定序列全长7519bp,与内源性绵羊肺腺瘤病毒代表株enJSRV-20核苷酸同源性为99．4％,与外源性绵羊肺腺瘤病毒代表株AF357971．1的核苷酸同源性为90．4％。基于全基因组序列核苷酸的系统进化发生树分析结果显示,扩增序列与enJSRV-20的亲缘关系最近。 相似文献
5.
绵羊肺腺瘤病毒中国NM株gag基因3''''端951 bp段的分子克隆与序列分析 总被引：4，自引：1，他引：3
究参照Genebank中已发表的绵羊肺腺瘤病毒的全基因序列,设计合成一对引物,对JSRV中国NM株的gag基因3'端中主要编码核衣壳(NC)蛋白的基因段进行PCR扩增,产物经琼脂糖凝胶电泳分析,呈现一条约951 bp的特异条带,将其回收后克隆人pMD-18T载体中,并进行序列测定.结果表明,与南非代表株(基因序列号NC-001494)的gag基因序列比较,核苷酸同源性为95.4%,推导出的氨基酸同源性为95%.与美国代表株(基因序列号AF105220)的gag基因序列比较,核苷酸同源性为91.3%,氨基酸同源性为91%.这是我国首次报道的绵羊肺腺瘤病毒的gag基因的一段序列,为我国科研工作者进行更深入的研究奠定基础. 相似文献
6.
绵羊肺腺瘤病毒NM株前病毒gag基因的克隆与序列分析 总被引：4，自引：0，他引：4
参照GenBank中已发表的绵羊肺腺瘤病毒(JSRV)的全基因序列，设计合成3对引物，对JSRVNM株的gag基因分3段进行PCR扩增，经琼脂糖凝胶电泳分析，分别呈3条531、888和949 bp的特异条带，将其分别克隆人pMD-18T载体中，进行序列测定并拼接序列，得到完整的gag基因序列。分析结果表明，与南非代表株(基因序列号NC-001494)的gag基因序列比较，核苷酸同源性为89．0%，推导出的氨基酸同源性为90％。与美国代表株(基因序列号AF105220)的gag基因序列比较，核苷酸同源性为86．3％，氨基酸同源性为87％。 相似文献
7.
用位于内源性禽白血病病毒ev/J 5′和3′长末端序列(LTR)上的引物,对我国的灵山麻鸡、江汉鸡、肉鸡和SPF鸡正常基因组中内源性ev/J进行扩增,结果扩增出长约为4 kb、2.2 kb和2 kb 3种大小的片段。测序分析表明,这些片段分属于Ⅱ型和Ⅳ型ev/J以及另一类具有更大缺失的ev/J基因。ev/J在这几个品系鸡中分布并不完全相同,肉鸡和SPF鸡均具有Ⅱ型和Ⅳ型ev/J,江汉鸡只含Ⅱ型ev/J,而灵山麻鸡则含有另一类具有更大缺失的ev/J基因。此次获得的Ⅳ型ev/J序列不仅相互之间高度同源,而且其env基因与外源性ALV-J原型株HPRS-103env基因以及我国外源性ALV-J主要分离株env基因也高度同源,表明内源性ev/J在宿主正常基因组中高度保守。 相似文献
8.
《动物医学进展》2015,(10)
为了解犬副流感病毒(CPIV)的基因组结构与遗传进化特征,应用RT-PCR分段扩增HRB-V毒株的5个片段,f1、f2、f3以及5′-末端和3′-末端,克隆到pHW2000载体中并进行测序。经序列拼接获得全基因组序列,与GenBank登录的副流感病毒代表毒株进行对比分析。基因序列分析表明,HRB-V株基因组全长15 246nt,含5′-末端、3′-末端和7个ORF具有副流感病毒基因组结构的典型特征;将结构蛋白F、HN以及全基因序列与参考毒株进行比较分析并做遗传进化树,结果表明,HRB-V与CPI-和CPI+属同一分支,亲缘性相近。核苷酸以及氨基酸序列同源性分析结果表明,HRB-V与各病毒株的F基因的核苷酸同源性在96.5%~98.8%之间,氨基酸同源性为94.2%~98%;HN基因的核苷酸同源性在98.1%~99.5%之间,氨基酸同源性为97%~99.3%。以上试验证明副流感病毒人、猴、猪和犬SV5分离株基因差异较小,具有高度同源性。研究副流感病毒分子克隆的结构,为研究犬副流感病毒反向遗传操作技术平台奠定基础。 相似文献
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10.
绵羊肺腺瘤病毒NM株囊膜基因的定向克隆与融合表达 总被引：1，自引：0，他引：1
根据绵羊肺腺瘤病毒(SJRV)NM株基因组全序列，设计了1对包含囊膜(env)基因在内的引物，并在5′端分别设计了HindⅢ和BnmHⅠ酶切位点，利用PCR技术扩增env基因，通过连接反应将其克隆于pET-30b表达载体上。序列分析表明，env基因全长1836bp，编码612个氨基酸残基；构建表达重组质粒env—pET—30b，转化大肠埃希氏菌BL21，经IPTG于37℃诱导培养，SDS—PAGE电泳分析表明，表达的env基因融合蛋白分子质量约为69ku。 相似文献
11.
绵羊基因组中内源性绵羊肺腺瘤病毒相关序列的确定 总被引：1，自引：0，他引：1
绵羊都含有与绵羊肺腺瘤病毒(Jaagsiekte sheep retrovirus,JSRV)密切相关的15~20拷贝内源性绵羊肺腺瘤病毒(enJSRV)相关序列。宿主可利用内源性病毒来预防致病性反转录病毒的感染,一些内源性病毒可以有效地干扰相关外源性病毒的复制。本试验通过分子生物学手段确定了蒙古绵羊基因组中含有enJSRV6和enJSRV10两个内源性病毒基因而内蒙古白绒山羊中未发现。通过比较内、外源病毒LTR序列的酶切图谱,获得专一作用外源性病毒的核酸内切酶M spⅠ、TfiⅠ、BsaWⅠ,如果将酶切与聚合酶链式反应(PCR)相结合,不需要经过测序就可分辨enJS-RV和外源性绵羊肺腺瘤病毒(exJSRV),形成"酶切-PCR"检测技术,将为绵羊肺腺瘤病的快速诊断提供了新的手段。 相似文献
12.
Maeda N Inoshima Y Oouchi S Uede T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2011,73(11):1493-1495
Surveillance of jaagsiekte sheep retrovirus (JSRV) infection was performed by polymerase chain reaction (PCR) of blood DNA samples collected from 40 sheep and goats in 10 different flocks in Hokkaido, the northern island of Japan. No exogenous (oncogenic) JSRV sequence was detected by PCR in these samples, while the ovine endogenous retrovirus sequence was successfully amplified in all samples. Our paper is the first demonstration of JSRV surveillance in Japan and shows no evidence of oncogenic JSRV infection in sheep and goats in Hokkaido. 相似文献
13.
Ovine pulmonary adenocarcinoma is caused by jaagsiekte sheep retrovirus. To gain insight into the histogenesis and viral pathogenesis of this neoplasm, the tumor cell phenotypes and differentiation state were correlated with the distribution of jaagsiekte sheep retrovirus capsid protein in neoplastic and normal cells of the lung in nine naturally occurring and 12 experimentally induced cases of ovine pulmonary adenocarcinoma. Overall, 82% of tumor cells had ultrastructural features consistent with alveolar type II cells, 7% of tumor cells had features of Clara cells, and 11% of tumor cells were insufficiently differentiated to classify. The proportion of the neoplastic cell phenotypes varied within tumors, and no tumor consisted of a morphologically uniform cell population. To further characterize the neoplastic cell population, sections of tumors were immunostained with antibodies to surfactant protein A, surfactant protein C, and Clara cell 10-kd protein. Overall, surfactant proteins A and C were expressed in 70% and 80% of tumor cells, respectively, whereas Clara cell 10-kd protein was expressed in 17% of tumor cells. Jaagsiekte sheep retrovirus capsid protein was detected in 71% of tumor cells and in macrophages (5/21 tumors examined) and in nonneoplastic alveolar and bronchiolar cells (6/14 tumors). Expression of this viral protein in neoplastic cells, classified morphologically and by immunophenotyping primarily as of the alveolar type II lineage, implies an important role for specific virus-cell interactions in the pathogenesis of ovine pulmonary adenocarcinoma. 相似文献
14.
利用ALV-J gp85基因两侧的序列片段为引物,从山东潍坊某肉种鸡场送检的7日龄肉种鸡基因组中完整地扩增了内源性类ALV-J gp85基因。利用ALV-J env基因特异性引物不能扩增出长度为2.2kb的目的片段,用ALV-J特异性单抗JE-9做间接免疫荧光检测,也呈现阴性反应。这种内源性类ALV-J gp85基因与已报道的禽内源性反录病毒EAV-HP序列的同源性为85.6%～93.8%,其中与ADOL 0系来航鸡EAV-HP的相似性最高(93.8%)。所得到的内源性类ALV-J gp85基因与外源性ALV-J毒株HPRS-103、SD07LK1、NX0101的同源性分别为93.8%、91.8%和91.7%。并且与外源性ALV-J gp85基因具有相似的Jameson-Worlf抗原表位优势。这种内源性类gp85基因很可能与肉雏鸡早期感染ALV-J后所呈现的免疫耐受现象有关。 相似文献
15.
Stewart H Jarrett O Hosie MJ Willett BJ 《Veterinary immunology and immunopathology》2011,143(3-4):325-331
Full length endogenous feline leukemia virus (FeLV) proviruses exist within the genomes of many breeds of domestic cat raising the possibility that they may also exist in a transmissible exogenous form. Such viruses would share receptor usage with the recombinant FeLV-B subgroup, a viral subgroup that arises in vivo by recombination between exogenous subgroup A virus (FeLV-A) and endogenous FeLV. Accordingly, all isolates of FeLV-B made to date have contained a "helper" FeLV-A, consistent with their recombinatorial origin. In order to assess whether endogenous viruses are transmitted between cats, we examined primary isolates of FeLV for which the viral subgroup had been determined for the presence of a subgroup B virus that lacked an FeLV-A. Here we describe the identification of two primary field isolates of FeLV (2518 and 4314) that appeared to contain subgroup B virus only by classical interference assays, raising the possibility of between-host transmission of endogenous FeLV. Sequencing of the env gene and U3 region of the 3' long terminal repeat (LTR) confirmed that both viral genomes contained endogenous viral env genes. However the viral 3' LTRs appeared exogenous in origin with a putative 3' recombination breakpoint residing at the 3' end of the env gene. Further, the FeLV-2518 virions also co-packaged a truncated FeLV-A genome containing a defective env gene, termed FeLV-2518(A) whilst no helper subgroup A viral genome was detected in virions of FeLV-4314. The acquisition of an exogenous LTR by the endogenous FeLV in 4314 may have allowed a recombinant FeLV variant to outgrow an exogenous FeLV-A virus that was presumably present during first infection. Given time, a similar evolution may also occur within the 2518 isolate. The data suggest that endogenous FeLVs may be mobilised by acquisition of exogenous LTRs yielding novel viruses that type biologically as FeLV-B. 相似文献
16.
猪内源性反转录病毒（PERV）是与猪-人异种移植病原安全性密切相关的一类病毒。env基因编码病毒的囊膜蛋白,它与病毒的亚型分类、宿主感染范围、细胞的嗜性以及对宿主细胞的感染机制、诱导宿主产生中和抗体等密切相关。本研究利用RT-PCR的方法,从五指山小型猪外周血淋巴细胞中扩增PERV的囊膜蛋白基因并进行测序,随后用生物信息学相关软件和方法,对PERV-Env蛋白二级结构及B细胞表位进行预测。经综合分析评价,结果发现PERV-Env蛋白有18个可能的B细胞优势抗原表位区域,7个可能的糖基化位点。该分析预测结果不但有利于PERV疫苗的设计、单抗及诊断试剂研制,而且将有助于分析Env蛋白的功能及PERV对人源细胞的感染机制。 相似文献
17.
Ramírez H Glaria I de Andrés X Martínez HA Hernández MM Reina R Iráizoz E Crespo H Berriatua E Vázquez J Amorena B de Andrés D 《Veterinary journal (London, England : 1997)》2011,190(1):169-172
Nucleotide sequences of small ruminant lentiviruses (SRLVs) were determined in sheep and goats, including progeny of imported animals, on a farm in Mexico. On the basis of gag-pol, pol, env and LTR sequences, SRLVs were assigned to the B1 subgroup, which comprises caprine arthritis-encephalitis virus (CAEV)-like prototype sequences mainly from goats. In comparison with CAEV-like env sequences of American and French origin, two putative recombination events were identified within the V3-V4 and V4-V5 regions of the env gene of a full length SRLV sequence (FESC-752) derived from a goat on the farm. 相似文献
18.
A Payne D F York E M de Villiers D W Verwoerd G Quérat V Barban N Sauze R Vigne 《The Onderstepoort journal of veterinary research》1986,53(1):55-62
In the course of attempts to grow the jaagsiekte retrovirus in cell culture, a typical lentivirus was isolated for the first time in South Africa from adenomatous lungs. Morphologically the virus could not be distinguished from other lentiviruses, but serologically it was shown to be more closely related to visna virus than to caprine arthritis-encephalitis virus. However, a preliminary restriction enzyme analysis of the linear proviral DNA of this new lentivirus (SA-DMVV) revealed that it is significantly district from visna virus and CAEV and therefore may represent a third type of lentivirus. Antibodies to the virus were demonstrated in a number of sheep in various parts of the country, but a direct link to a disease condition was not found. Attempts to produce lung lesions by intratracheal injection of the virus have been unsuccessful to date but a transient arthritis was produced by intraarticular inoculation. Viral replication seems to be enhanced in jaagsiekte lungs. 相似文献
19.
S Pr?sch 《Archiv fuer experimentelle veterinaermedizin》1990,44(6):935-939
RD114 virus is an endogenous xenotropic Type C retrovirus of domestic cat. Previously, it had been shown that genomic DNA of cat contained approximately 20 copies of RD114-related sequences. Only one encoded for the replication-competent RD114 virus. The endogenous sequences exhibited substantial sequence conservation within the gag and pol genes and LTRs, but were characterized by deletions and substitutions within the env region. The endogenous cat retroviral element ECE2 was isolated by screening a genomic DNA library of cat liver DNA with an env-specific cDNA of RD114 virus. It contained an env region which differed from all RD114-related sequences so far isolated and was homologous to the corresponding region of replication-competent RD114 with regard to their restriction map and partial sequence analysis (p20). Otherwise, ECE2 had a deletion of approximately 1 kbp in the putative pol gene and, therefore, did not represent the locus of inducible RD114 virus. 相似文献