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K亚群禽白血病病毒gp85单因子血清的制备及其特异性鉴定 被引量:3
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作者 孙鹏 陈孜孟 +4 位作者 赵国梁 崔宁 祖铭 崔治中 苏帅 《中国畜牧兽医》 CAS 北大核心 2016年第2期371-376,共6页
为了制备一种快速特异性识别K亚群禽白血病病毒(ALV-K)的单因子血清,本试验将ALV-K接种DF1细胞,提取感染细胞DNA作为模板,PCR扩增1 005bp ALV-K-gp85和510bp ALV-K-gp85(fragment)基因。将其正确的阅读框插入表达载体pET-28a(+)中,转化... 为了制备一种快速特异性识别K亚群禽白血病病毒(ALV-K)的单因子血清,本试验将ALV-K接种DF1细胞,提取感染细胞DNA作为模板,PCR扩增1 005bp ALV-K-gp85和510bp ALV-K-gp85(fragment)基因。将其正确的阅读框插入表达载体pET-28a(+)中,转化大肠杆菌BL21(Rosetta)菌株中表达。将表达蛋白常规免疫昆明白小鼠制备抗血清。结果表明成功获得39.85和22.01ku的重组融合蛋白,且具有良好的免疫原性。间接免疫荧光试验(IFA)结果表明两血清均可与ALV-K反应,而不与ALV-A、ALV-B和ALV-J反应。本试验首次研制获得能特异性识别ALV-K的单因子血清,为K亚群禽白血病的快速检测奠定了基础。 展开更多
关键词 K亚群禽白血病病毒 GP85基因 表达 间接免疫荧光试验(IFA) 单因子血清
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Isolation of Three Strains of Avian Leukosis Virus Subgroup J and gp85 Gene Sequence Analysis
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作者 LIU Li-na LUO Qing-ping +5 位作者 CHENG Guo-fu HU Xue-ying YANG Jun ZHANG Lin SHAO Hua-bin LI Shao-wen 《Animal Husbandry and Feed Science》 CAS 2013年第1期11-13,52,共4页
[Objective] To isolate three strains of avian leukosis virus subgroup J(ALV -J) , and then amplify and sequence the gp85 gene. [ Method] Three strains of ALV- J were isolated from Hubei Province, which were identifi... [Objective] To isolate three strains of avian leukosis virus subgroup J(ALV -J) , and then amplify and sequence the gp85 gene. [ Method] Three strains of ALV- J were isolated from Hubei Province, which were identified by pathological anatomy, DF- 1 cell culture and RT- PCR. And then they were named HB1002, HB1003 and HB1009, respectively. [ Result] Test sequence analysis showed, the length of gp85 gene was 921 bp, consistent with expect result; the nucleotide homology between the three isolates was in 97.7% -99.7% ,the homology of amino acid was in 95.1% - 99%. The nucleotide homology between HPRS - 103 and the three isolates was in the 94.1% - 94.8% ; and the nucleotide homolo- gy between other ALV -J and the three isolates was in 87.6% -97.3%. The phylogenetic trees analysis showed that the homology of JS09GY6 vi- rus and the three isolates was nearest, in 95.2% -97.3%. [ Conclusion] In the test, the three strains of virus which were isolated were ALV- J. 展开更多
关键词 ALV - J gp85 gene Sequence analysis
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Expression of Endogenous Retrovirus ev/J gp85 Gene and Analysis of Its Immunoreactivity in Comparison with Exogenous Viral Protein
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作者 Yu-ying YANG Ai-jian QIN +1 位作者 Xiong-yan LIANG Shu-mei TONG 《Virologica Sinica》 SCIE CAS CSCD 2008年第5期369-377,共9页
The envelope gene gp85 of ev/J, a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian leukosis virus. This repor... The envelope gene gp85 of ev/J, a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian leukosis virus. This report described expression of ev/J envelope gene gp85 derived from commercial meat-type chicken using the Invitrogen Bac-to-Bac baculovirus expression system. The antigenicity and immunoreactivity of the recombinant endogenous gp85 gene product (SU) were analyzed by indirect immunofluorescence, Western blot, indirect and blocking Enzyme-Linked ImmunoSorbent Assay (ELISA) using JE9 monoclonal antibody (MAb) against the envelope protein of ALV-J (ADOL-4817), positive mouse antiserum against the ev/J gp85 SU and sera from chicken naturally infected with ALV-J. The results showed that the ev/J gp85 SU can bind specifically to JE9 MAb and antiserum from chicken naturally infected with ALV-J, and the binding reactivity between exogenous ALV-J gp85 SU and natural positive chicken serum against exogenous ALV-J can be blocked by positive mouse serum against the ev/J gp85 SU. It is concluded that recombinant endogenous gp85 gene product (SU) has close immunological relatedness to the envelope protein of exogenous ALV-J (ADOL-4817 and IMC10200 strain). 展开更多
关键词 Avian endogenous retrovirus ev/J Envelope gene gp85 EXPRESSION IMMUNOREACTIVITY
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禽白血病A亚群病毒gp85的单因子血清制备及其特异性鉴定 被引量:13
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作者 张恒 李传龙 +1 位作者 杨明 崔治中 《微生物学报》 CAS CSCD 北大核心 2011年第1期134-140,共7页
【目的】为了研究出一种能够针对A亚群禽白血病的快速特异性诊断试剂。【方法】将A亚群禽白血病病毒(ALV-A)SDAU09E1株接种于DF1细胞上,以感染细胞DNA为模板,通过PCR方法扩增出1023bp的ALV-A-gp85基因。将其正确阅读框架插入表达载体PET... 【目的】为了研究出一种能够针对A亚群禽白血病的快速特异性诊断试剂。【方法】将A亚群禽白血病病毒(ALV-A)SDAU09E1株接种于DF1细胞上,以感染细胞DNA为模板,通过PCR方法扩增出1023bp的ALV-A-gp85基因。将其正确阅读框架插入表达载体PET-32a(+)中,实现在BL21(Rosetta)宿主菌中表达。将纯化的融合蛋白常规免疫小鼠,制备得抗血清。【结果】实验成功获得52.8kDa的重组融合蛋白,且具有良好的免疫原性。间接免疫荧光试验(IFA)表明该血清可与ALV-A和ALV-B反应,但不与ALV-J反应。【结论】该实验首次在国内外研制出能用于鉴别性检测经典的A/B亚群ALV的单因子血清,可与ALV-J特异性单抗互补作用于外源性ALV感染的鉴别性诊断。我国鸡群同时受经典的ALV-A/B和新出现的ALV-J困扰,鉴别诊断非常必要,研究这种试剂具有较高的实用价值。 展开更多
关键词 A亚群禽白血病病毒 GP85基因 表达 IFA 单因子血清
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