| 麻鸭白细胞介素17的原核表达及鉴定 |
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| 引用本文： | 赵雯,潘志明,耿士忠,康喜龙,方强,丛秋霞,焦新安.麻鸭白细胞介素17的原核表达及鉴定[J].中国预防兽医学报,2011,33(5). |
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| 作者姓名： | 赵雯 潘志明 耿士忠 康喜龙 方强 丛秋霞 焦新安 |
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| 作者单位： | 扬州大学江苏省人兽共患病学重点实验室,江苏,扬州,225009 |
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| 基金项目： | 国家自然科学基金资助项目，江苏省自然科学基金资助项目，江苏省高校"青蓝工程"资助项目，扬州大学中青年学术带头人培养人选资助项目 |
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| 摘 要： | 为表达麻鸭白介素17(duIL-17),本研究提取ConA刺激的麻鸭脾脏淋巴细胞总RNA,RT-PCR扩增duIL-17基因片段,将该片段插入克隆载体pCR2.1中构建重组质粒pCR2.1-duIL17.将duIL-17基因片段亚克隆到原核表达载体pGEX-6P-1中,构建重组质粒pGEX-duIL17,将其转化到大肠杆菌BL21中,IPTG诱导表达.测序结果显示,duIL-17与参考序列(EU366165.1)相比较第174位碱基由C突变为T,并且为沉默突变.SDS-PAGE和western blot分析显示,诱导表达的重组蛋白GST-duIL17主要以包涵体形式存在,分子量大小约为45 ku,duIL-17与抗鸡IL-17抗体具有良好反应性.本研究为制备duIL-17单克隆抗体提供了实验依据.
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| 关 键 词： | 麻鸭 IL-17 原核表达 免疫反应性 |
Expression of spotted duck interleukin-17 gene in E.coli |
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| ZHAO Wen,PAN Zhi-ming,GENG Shi-zhong,KANG Xi-long,FANG Qiang,CONG Qiu-xia,JIAO Xin-an.Expression of spotted duck interleukin-17 gene in E.coli[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(5). |
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| Authors: | ZHAO Wen PAN Zhi-ming GENG Shi-zhong KANG Xi-long FANG Qiang CONG Qiu-xia JIAO Xin-an |
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| Institution: | ZHAO Wen,PAN Zhi-ming,GENG Shi-zhong,KANG Xi-long,FANG Qiang,CONG Qiu-xia,JIAO Xin-an (JiangSu Key Laboratory of Zoonosis,Yangzhou University,Yangzhou 225009,China) |
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| Abstract: | The duck interleukin-17 (duIL-17) gene was amplified by RT-PCR from the total RNA of conconavalin A (ConA)-stimulated spotted duck splenic lymphocytes. The gene was cloned into vector pCR2.1 to construct recombinant plasmid pCR2.1-duIL17. Sequencing result showed that duIL-17 had a silent mutation at 174 set from C to T, compared with the reference gene (EU366165.1). Then duIL-17 gene fragment was subcloned into the expression vector pGEX-6P-1, the resultant recombinant plasmid was transformed into E. coli ... |
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| Keywords: | spotted duck interleukin-17 prokaryotic expression immunoreactivity |
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