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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (12): 30-33.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

香蕉果实凝集素基因的克隆及原核表达

陈石,李春雨,孙清明,匡石滋,杨国顺,易干军   

  • 收稿日期:2009-01-16 修回日期:2009-02-11 出版日期:2009-06-20 发布日期:2009-06-20

Cloning and Expressing BanLec Gene from Pulp in Escherichia coli

Chen Shi,, Li Chunyu, Sun Qingming, Kuang Shizi, Yang Guoshun, Yi Ganjun   

  • Received:2009-01-16 Revised:2009-02-11 Online:2009-06-20 Published:2009-06-20

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摘要： 【研究目的】植物凝集素具有重要的生理功能，通过原核表达香蕉凝集素基因(BanLec)并探讨其功能具有重要意义。【方法】提取了巴西蕉果肉总RNA，利用RT-PCR方法扩增BanLec cDNA 全长序列，并对该序列进行分析。将BanLec克隆至表达载体pET-30a，并将筛选到的重组质粒转化大肠杆菌BL21(DE3)菌株中进行诱导表达。【结果】克隆到的BanLec cDNA 序列为460bp，开放读码框为426bp，编码142个氨基酸。与其它已知的香蕉凝集素基因相比较，核苷酸和氨基酸序列同源性分别为94%和93%以上。SDS-PAGE 分析结果表明，经IPTG诱导，BanLec基因以融合蛋白形式表达，相对分子量约为20kD。【结论】该研究为探讨香蕉凝集素的活性及生化功能等奠定基础。

关键词: 农业科技示范园, 农业科技示范园, 规划, 可持续发展, 山东泰山

Abstract: 【Objective】Plant lectin has important in vivo physiological functions, and it is necessary to expressing BanLec gene in Escherichia coli and investigate its functions. 【Method】Total RNA was extracted from Brazile banana pulp, and the BanLec cDNA sequence was amplified with Reverse Transcription Polymerse Chain Reaction (RT-PCR) and the sequence was analyzed. BanLec was cloned into pET-30a vector, and the recombinant plasmid pET-BanLec was screened. After transforming into Escherichia coli BL21(DE3), the protein was induced to express. 【Result】The Sequence was analyzed with DNAstar software and the result showed that its open reading frame was 426bp and encoded a protein with 142 amino acids. The alignment showed that this gene shared high similarities with other known BanLec genes on the levels of nucleotides and amino acids (94% and 93%, respectively). The result of SDS-PAGE demonstrated that the BanLec gene was expressed by induction and the expression products migrated at a size of 20kD. 【Conclusion】This research is helpful for investigating the activities or other physiological functions of banana lectins.