| 大豆rbcL基因克隆、序列分析及原核表达 |
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| 引用本文： | 刘晓庆,崔喜艳,丁志鑫,李海燕.大豆rbcL基因克隆、序列分析及原核表达[J].中国油料作物学报,2011,33(3):226-230. |
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| 作者姓名： | 刘晓庆 崔喜艳 丁志鑫 李海燕 |
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| 作者单位： | （吉林农业大学生命科学学院，吉林，长春130118）
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| 基金项目： | 国家转基因生物新品种培育重大专项，国家自然科学基金，吉林省教育厅吉教科合字[2009]第61号 |
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| 摘 要： | 利用叶绿体基因组保守性的特征，根据菜豆、豌豆、烟草的rbcL基因序列设计引物，从大豆叶绿体DNA中克隆rbcL基因，全长序列为1488bp，包括1449bp的开放阅读框，编码482个氨基酸。相似性比较显示，此序列与其它10个物种rbcL基因核苷酸的同源性为85.37%～95.31%，氨基酸的同源性为90.87%～96.47%。将该基因与表达载体pET-30a（+）连接，转化大肠杆菌Rosseta感受态细胞，PCR和酶切鉴定筛选阳性克隆，阳性菌液IPTG诱导后经10%SDS-PAGE分析，结果显示：诱导表达出分子量约为60kD的特异融合蛋白。
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| 关 键 词： | 大豆，rbcL基因，基因克隆，原核表达 |
Cloning, sequence analysis and prokaryotic expression of rbcL gene from Glycine max |
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| LIU Xiao-qing,CUI Xi-yan,DING Zhi-xin,LI Hai-yan.Cloning, sequence analysis and prokaryotic expression of rbcL gene from Glycine max[J].Chinese Journal of Oil Crop Sciences,2011,33(3):226-230. |
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| Authors: | LIU Xiao-qing CUI Xi-yan DING Zhi-xin LI Hai-yan |
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| Institution: | LIU Xiao-qing,CUI Xi-yan,DING Zhi-xin,LI Hai-yan(College of Life Sciences,Jilin Agricultural University,Changchun 130118,China) |
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| Abstract: | Rubisco is the key enzyme in plant photosynthesis.In this research,the large subunit of Rubisco,rbcL gene,from chloroplast of Glycine max was cloned and sequenced.Sequencing analysis indicated that this fragment was 1 488bp in size,including an ORF of 1 446bp,encoding a putative protein consisting of 482 amino acids.DNAman analysis results showed that the homologies of this gene with other species were from 85% to 96%,and the homologous amino acid sequences were from 90% to 97%.The rbcL gene was ligated int... |
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| Keywords: | Glycine max rbcL gene Gene cloning Prokaryotic expression |
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