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Cloning and expressions of G1-ITAM-like sequences in Hantaan virus by gateway technology
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 15
- DOI:
- 10.3321/j.issn:1000-2790.2007.15.001
- Key Word:
- 汉坦病毒;囊膜糖蛋白类;免疫受体酪氨酸活化基序
Abstract： 目的:克隆并表达汉滩病毒(HTNV)囊膜糖蛋白(G1)胞质区尾段ITAM样基序及其突变体.方法:采用RT-PCR方法,扩增HTNV G1蛋白ITAM样基序及其中2个保守酪氨酸残基突变基序编码基因,应用Gateway技术将目的基因克隆至pDESTTM15表达载体,在BL21-AI中经L-阿拉伯糖诱导表达,免疫印迹确证融合蛋白的表达.结果:成功地构建了2个含汉滩病毒G1胞质区尾段ITAM样基序编码序列表达载体pDEST- ITAM-L,pDEST-ITAM和1个含保守酪氨酸残基突变的G1 ITAM样基序编码序列表达载体pDEST-ITAM-YY-FF (Y615F,Y628F),并在大肠埃希杆菌中得到高效表达,免疫印迹证实目的蛋白与GST融合表达.结论:获得了3种与谷胱甘肽-S-转移酶(GST)融合的G1 ITAM样基序及其突变基序的重组蛋白, 为进一步探讨HTNV G1蛋白ITAM样基序在HFRS免疫信号传递中的作用奠定了基础.
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