| 狮头鹅PRL基因的克隆及原核表达 |
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| 引用本文： | 贾汝敏,吴慧英,刘铀,张丽.狮头鹅PRL基因的克隆及原核表达[J].畜牧兽医学报,2011,42(3). |
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| 作者姓名： | 贾汝敏 吴慧英 刘铀 张丽 |
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| 作者单位： | 1. 广东海洋大学动物科学系,湛江,524088
2. 广东海洋大学动物医学系,湛江,524088 |
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| 基金项目： | 湛江市科技招标项目(0609135); 广东海洋大学基金项目(C04095); 校企合作项目(0810314) |
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| 摘 要： | 为了进一步研究PRL对狮头鹅繁殖性能的生理调节机制,采用RT-PCR方法从狮头鹅脑垂体组织扩增获得PRL基因的cDNA序列,克隆至pMD18-T载体获得重组质粒pMD18-PRL,并进行序列测定和分析。将测序正确的cDNA序列定向克隆到pET32a(+),构建表达载体pET32a-PRL,并转化至BL21(DE3)大肠杆菌表达目的蛋白,经IPTG诱导后进行SDS-PAGE检测和Western Blot分析。狮头鹅PRL基因编码区含有600个核苷酸,编码199个氨基酸(GenBank No.GQ856665),与其它禽类PRL具有高度保守性;狮头鹅PRL蛋白二级结构由多个α螺旋和β转角及无规卷曲构成,推测N端70～76、95～102、150～155和207～213区段为其抗原表位的优势区。SDS-PAGE检测表明狮头鹅PRL基因在大肠杆菌中获得了高效表达,可溶性表达产物约占全菌总蛋白的53.6%,融合蛋白分子量约41ku,并用镍柱亲和层析法分离纯化融合蛋白。Western Blot分析证实了所获的融合蛋白具有较强抗原性。试验结果为进一步研究PRL基因的生物功能及其对狮头鹅就巢、产蛋等生产性状的影响奠定了基础。
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| 关 键 词： | 狮头鹅 催乳素 克隆 基因表达 PRL蛋白 |
Cloning and Prokaryotic Expression of PRL Gene in Shitou Goose |
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| JIA Ru-min,WU Hui-ying,LIU You,ZHANG Li.Cloning and Prokaryotic Expression of PRL Gene in Shitou Goose[J].Acta Veterinaria et Zootechnica Sinica,2011,42(3). |
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| Authors: | JIA Ru-min WU Hui-ying LIU You ZHANG Li |
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| Institution: | JIA Ru-min1,WU Hui-ying1,LIU You2,ZHANG Li1 (1.Department of Animal Science,Guangdong Ocean University,Zhanjiang 524088,China,2.Department of Veterinary,China) |
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| Abstract: | To further study the physiological regulatory mechanisms of PRL on the reproductive performance of Shitou goose.The cDNA of goose prolactin(PRL) gene was amplified from anterior pituitary gland of Shitou goose by RT-PCR.Then the PCR product was cloned into the pMD18-T vector to construct the pMD18-PRL for sequencing.And then the cDNA was subcloned into the prokaryotic expressing vector pET32a(+).Subsequently,the recombinant plasmid pET32-PRL was transformed into the E.coli BL21(DE3) expression bacteria,then... |
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| Keywords: | Shitou goose prolactin cloning gene expression PRL protein |
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