XUE Xiao-Beng, XU Zhi-Kai, MA Wen-Yu, YIN Wen, ZHANG Fang-Lin, YAN Yan, TUN Xin-An and BAI Wen-Chao. Expression of Truncated HTNV Nncleoprotein and Analysis of Antigenic epitope[J]. Virologica Sinica, 2000, 15(3): 220-225.
Citation: XUE Xiao-Beng, XU Zhi-Kai, MA Wen-Yu, YIN Wen, ZHANG Fang-Lin, YAN Yan, TUN Xin-An, BAI Wen-Chao. Expression of Truncated HTNV Nncleoprotein and Analysis of Antigenic epitope .VIROLOGICA SINICA, 2000, 15(3) : 220-225.

汉滩病毒核蛋白的分段表达及抗原表位分析

  • 在大肠杆菌中对汉滩病毒S基因 4种不同长度片段的重组表达质粒进行诱导表达。结果表明表达的 4种GST NP融合蛋白均以不溶性包含体形式存在于菌体细胞内 ,表达量分别占菌体蛋白总量的 2 9- 36 % ,分子量分别约为 72kD、6 6kD、54kD和 4 4kD。Westernblot显示 54kD和 72kD融合蛋白用酶标抗汉滩病毒NPMcAb 1A8和抗GSTMcAb 3C11染色呈阳性反应。 6 6kD和4 4kD融合蛋白仅与酶标 3C11呈阳性反应。用凝血酶切去GST ,获得 4种汉滩病毒重组核蛋白(rNP) ,分子量分别约为 4 4kD、4 0kD、2 6kD和 16kD。用 19株McAb对表达产物做抗原位点分析 ,结果完整的rNP可与 19株McAb中的 13株反应 ,McAb反应谱与天然NP相同 ;S1.1kb表达产物 (N 端 1 37和C端 4 0 2 4 2 9位aa缺失 )和S 0 .5kb表达产物 (N 端 1 2 74位aa缺失 )与 19株McAb均不发生反应 ;S0 .7kb表达产物 (C 端 2 75 4 2 9位aa缺失 )可与 5株组特异性McAb反应。表明汉坦病毒核蛋白上的抗原位点主要存在于N 端 1 37位aa区段内。

Expression of Truncated HTNV Nncleoprotein and Analysis of Antigenic epitope

  • The expressing vector carring various truncated fragments of S gene of HTNV strain 76 118 was constructed and the vector efficienly expressed in E.coli . The result demonstrated that the GST NP fusion proteins exist in the form of inclusion bodies in E.coli , the expressing amount accounted for 29 36% of the total proteins, and the molecular weights were 72 kD, 66 kD, 54 kD and 44 kD, respectively. Western blot showed that all of the four fusion proteins were positive stained with HPR labeled anti GST McAb 3C11, but only the 72 kD and 54 kD fusion proteins were positive stained with HRP labeled anti NP McAb 1A8. Four recombinant NP (rNP), which molecular weights were 44 kD, 40 kD, 26 kD and 16 kD respectively, were obtained by removing GST from purified GST NP fusion proteins with thrombin. Mapping of antigenic epitope was done by 19 strains McAb. The result showed that 72 kD fusion protein could react with 13 strains McAb, which was same as authentic NP of HTNV. The truncated fusion protein (d

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    Expression of Truncated HTNV Nncleoprotein and Analysis of Antigenic epitope

    • 1. Dept.of Microbiology,Fourth Military Medical University,xi'an 4 710032,China

    Abstract: The expressing vector carring various truncated fragments of S gene of HTNV strain 76 118 was constructed and the vector efficienly expressed in E.coli . The result demonstrated that the GST NP fusion proteins exist in the form of inclusion bodies in E.coli , the expressing amount accounted for 29 36% of the total proteins, and the molecular weights were 72 kD, 66 kD, 54 kD and 44 kD, respectively. Western blot showed that all of the four fusion proteins were positive stained with HPR labeled anti GST McAb 3C11, but only the 72 kD and 54 kD fusion proteins were positive stained with HRP labeled anti NP McAb 1A8. Four recombinant NP (rNP), which molecular weights were 44 kD, 40 kD, 26 kD and 16 kD respectively, were obtained by removing GST from purified GST NP fusion proteins with thrombin. Mapping of antigenic epitope was done by 19 strains McAb. The result showed that 72 kD fusion protein could react with 13 strains McAb, which was same as authentic NP of HTNV. The truncated fusion protein (d

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