| 集胞藻PCC6803基因sll0853的克隆、优化表达及其结构功能预测 |
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| 引用本文： | 刘欣,陈思礼,梅菊,尤隽丹.集胞藻PCC6803基因sll0853的克隆、优化表达及其结构功能预测[J].山西大学学报(自然科学版),2012(3):552-556. |
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| 作者姓名： | 刘欣 陈思礼 梅菊 尤隽丹 |
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| 作者单位： | 中南民族大学生命科学学院 |
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| 基金项目： | 国家自然科学基金(31001099/C190101);湖北省全球基金艾滋病项目(SZY10008);微生物与生物转化中南民族大学重点实验室项目(XJS09002) |
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| 摘 要： | 从集胞藻(Synechocystis sp.)PCC6803提取总DNA,利用PCR扩增目的基因sll0853,构建重组T-0853克隆载体和pET-0853原核表达载体.为了提高sll0853在大肠杆菌E.coli BL21(DE3)中的表达量,通过改变诱导温度、诱导时间及诱导剂浓度等条件对表达量产生影响,以SDS-PAGE电泳分析证明sll0853基因蛋白表达的最佳条件.结果表明:目的蛋白在28℃、0.2mmol/L IPTG、诱导6h表达量分别达高峰.通过生物信息学软件预测基因sll0853可能具有裂合酶的功能.
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| 关 键 词： | 集胞藻PCC6803 sll0853 克隆表达 结构功能预测 |
Cloning,Optimization of Prokaryotic Expression of sll0853 and Prediction of the Structure and Function |
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| LIU Xin,CHEN Si-li,MEI Jü,YOU Jun-dan.Cloning,Optimization of Prokaryotic Expression of sll0853 and Prediction of the Structure and Function[J].Journal of Shanxi University (Natural Science Edition),2012(3):552-556. |
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| Authors: | LIU Xin CHEN Si-li MEI Jü YOU Jun-dan |
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| Institution: | (College of Life Science,South-central University for Nationalities,Wuhan 430074,China) |
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| Abstract: | The total DNA was extracted from Synechocystis sp.PCC6803,and sll0853 gene was screened and amplified by PCR.The sll0853 gene was connected with cloning vector,pMD18-T and a prokaryotic expression vector,pET-30a.In order to improve the expression level of sll0853 in E.coli BL21(DE3),the optimal conditions for expression of fusion protein were analyzed by SDS-PAGE at different concentration,temperature and time of IPTG.The results showed that the fusion protein expression of recombinant plasmid in E.coli BL21(DE3) was the best at 0.2 mmol/L of IPTG for 6 h at 28 ℃.In addition,the protein of sll0853 maybe have nature of CpeS through the prediction of bioinformatics. |
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| Keywords: | Synechocystis sp PCC6803 sll0853 cloning and expression prediction of the structure and function |
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