| 长白猪甘露聚糖结合凝集素A基因的 克隆与原核表达 |
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| 作者姓名： | 姚 欣 刘玉芬 唐高霞 刘洪雨 |
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| 作者单位： | [1]哈尔滨师范大学生命科学与技术学院,哈尔滨150025 [2]黑龙江省动物卫生监督所,哈尔滨150095 |
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| 基金项目： | 哈尔滨师范大学校科研和教改项目，黑龙江省教育厅科学技术研究项目 |
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| 摘 要： | 从长白猪肝脏细胞中提取总RNA,通过RT-PCR法扩增pMBL-A基因,与pMD18-T载体连接进行TA克隆,得到猪甘露聚糖结合凝集素A(poreine mannan-binding lectin A,pMBL-A)基因.再亚克隆到pPROEXHTb表达载体上,构建重组表达质粒.将阳性重组表达质粒转化到大肠杆菌中诱导表达重组pMBL-A蛋白,通过SDS-PAGE检测蛋白的表达情况.结果成功扩增得到包括完整开放读码框长为875bp的全长cDNA片段,并原核表达了重组蛋白,为进一步研究该蛋白的遗传特征以及为猪遗传育种方面的研究提供依据.
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| 关 键 词： | 猪甘露聚糖结合凝集素A 基因克隆 载体构建 原核表达 |
Cloning and Prokaryotic Expression of Porcine Mannan-binding Lectin A Gene |
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| Authors: | Yao Xin Liu Yufen Tang Gaoxi Liu Hongyu |
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| Institution: | Life Science and Technology Department in Harbin Normal University, Harbin 150025;
Animal Public Health Department of Heilongjiang Province, Harbin 150095 |
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| Abstract: | Total RNA was extracted from liver cells of landrace, pMBL-A gene was amplified by RT-PCR. It was cloned to pMD18-T vector, then the mannan-binding lectin A gene of landrace was obtained. It was subcloned to pPROEXHTb vector for expression, recombinant expression plasmid was constructed. The recombinant expression plasmid was transformed into E.coli and was induced to express recombinant protein of pMBL-A. The expressed protein was identified by SDS-PAGE. The cDNA of pMBL-A gene was successfully cloned, its full-length was 875bp including the whole ORF. Recombination protein was pronuleus expressed, it provides affordance for the research of genetic character of the gene and genetic breeding of pigs. |
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| Keywords: | porcine mannan-binding lectin Azz gene cloningzz vector constructionzz prokaryotic expressionzz |
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