| 鸭DCN基因编码区扩增、序列分析及其在大肠杆菌中的表达、蛋白纯化 |
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| 引用本文： | 余海跃,刘贺贺,金海波,王浩瀚,陈华丽,施洋洋,陈禧,李亮,王继文.鸭DCN基因编码区扩增、序列分析及其在大肠杆菌中的表达、蛋白纯化[J].中国家禽,2012,34(8):12-16. |
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| 作者姓名： | 余海跃 刘贺贺 金海波 王浩瀚 陈华丽 施洋洋 陈禧 李亮 王继文 |
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| 作者单位： | 四川农业大学动物遗传育种研究所,四川雅安,625014 |
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| 基金项目： | 国家高技术研究发展计划项目 |
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| 摘 要： | 采用RT-PCR方法从鸭腿肌总RNA中扩增了鸭核心蛋白聚糖基因编码区序列,进行序列分析,并将编码鸭核心蛋白聚糖成熟蛋白的核酸片段连入原核表达载体pET-32a(+)中,加入IPTG诱导其表达后,亲和层析纯化表达的蛋白。采用SDS-PAGE检测,用质谱方法对表达的蛋白进行鉴定。研究成功克隆出鸭核心蛋白聚糖基因编码区序列,序列分析表明,鸭DCN编码区序列由1074个碱基组成,编码357个氨基酸,其中信号肽有16个氨基酸。将鸭DCN蛋白划分为9个结构域(LRR1-9),结合人DCN序列进行序列分析推测结构域中LRR4、5与TGF-β可能存在一定关系。SDS-PAGE检测和质谱鉴定结果表明获得了分子量为54.7ku的鸭DCN融合蛋白。
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| 关 键 词： | 鸭核心蛋白聚糖 CDS 原核表达 蛋白纯化 |
Amplification and Analysis of Duck DCN Gene CDS and Its Expression in Escherichia coil and Protein Purification |
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| YU Haiyue
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LIU Hehe
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JIN Haibo
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WANG Haohan
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CHEN Huali
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SHI Yangyang
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CHEN Xi
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LI Liang
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WANG Jiwen.Amplification and Analysis of Duck DCN Gene CDS and Its Expression in Escherichia coil and Protein Purification[J].China Poultry,2012,34(8):12-16. |
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| Authors: | YU Haiyue LIU Hehe JIN Haibo WANG Haohan CHEN Huali SHI Yangyang CHEN Xi LI Liang WANG Jiwen |
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| Institution: | (Institute of Animal Breeding & Genetic,Sichuan Agricultural University,Ya’an,Sichuan 625014) |
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| Abstract: | DCN coding domain sequence of duck was obtained from the total RNA of adult duck using RT-PCR method.Then the duck DCN mature peptide cDNA was inserted into pET-32a(+) vector and transformed into Escherichia coli Rosseta.And then it was induced by IPTG,the recombinant expression products were detected by SDS-PAGE and confirmed by mass spectrum.The results of sequencing analysis showed that the full duck DCN coding domain sequence was 1 074 bp,encoding for 357 amino acids,and the deduced CDS amino acid sequence contained a 16 amino acids putative signal peptide.The DCN amino acid sequence of duck was composed of nine domains,According to the report about human,there might be certain relationship between LRR4、5 and TGF-β.The recombinant expression products were purified by affinity chromatography.The molecular weight of DCN protein was 54.7 ku through SDS-PAGE and mass spectroscopy. |
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| Keywords: | decorin CDS prokaryotic expression protein purification |
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