| 猪视黄醇结合蛋白在毕赤酵母中的表达及检测 |
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| 引用本文： | 张冬杰,谷溪,杨国伟,刘娣. 猪视黄醇结合蛋白在毕赤酵母中的表达及检测[J]. 东北农业大学学报, 2009, 40(3) |
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| 作者姓名： | 张冬杰 谷溪 杨国伟 刘娣 |
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| 作者单位： | 黑龙江省农业科学院,哈尔滨,150086;黑龙江省农业科学院,哈尔滨,150086;黑龙江省农业科学院,哈尔滨,150086;黑龙江省农业科学院,哈尔滨,150086 |
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| 基金项目： | 中国农科院畜禽种质资源标准化整理整合及共享平台建设项目，黑龙江省博士后启动基金 |
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| 摘 要： | 采用RT-PCR技术从猪的肝脏中扩增到RBP基因,将其与巴斯德毕赤酵母分泌型表达载体pPICZαC重组构建了重组表达载体pPICZαC-RBP,测序正确后将其用SacⅠ内切酶线性化后采用电穿孔法转化巴斯德毕赤酵母菌GS115,经ZeocinTM抗性筛选高拷贝重组菌株后用甲醇诱导表达,Western Blot分析结果表明,诱导表达的培养上清液中表达出具有生物活性的RBP重组蛋白。
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| 关 键 词： | 猪视黄醇结合蛋白 毕赤酵母 真核表达 |
Expression and detection of RBP gene of pig in Pichia pastoris |
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| ZHANG Dongjie,GU Xi,YANG Guowei,LIU Di. Expression and detection of RBP gene of pig in Pichia pastoris[J]. Journal of Northeast Agricultural University, 2009, 40(3) |
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| Authors: | ZHANG Dongjie GU Xi YANG Guowei LIU Di |
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| Affiliation: | Heilongjiang Academy of Agricultural Sciences;Harbin 150086;China |
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| Abstract: | RBP gene was cloned from liver of pig using RT-PCR technology, and then was inserted into asecretory expression vector pPICZαC, the expression vector pPICZαC-RBP was constructed successfully. The recombinant was linearized with SacⅠ and electroporated into the yeast cell GS115. The multi-copy recombinant P. pastoris strains was screened by ZeocinTM and induced by methanol. The expressed product was analyzed by Western Blot tests. Results showed that RBP gene was expressed successfully in culture supernatant. |
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| Keywords: | pig retinol-binding protein Pichia pastoris eukaryotic expression |
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