| 优化感受态细胞制备方法提高转化效率的研究 |
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| 引用本文： | 王世伟,李旭业,张伟伟.优化感受态细胞制备方法提高转化效率的研究[J].齐齐哈尔大学学报(自然科学版),2009,25(2). |
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| 作者姓名： | 王世伟 李旭业 张伟伟 |
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| 作者单位： | 1. 齐齐哈尔大学,生命科学与工程学院,黑龙江,齐齐哈尔161006
2. 黑龙江省畜牧研究所,黑龙江,齐齐哈尔161005 |
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| 基金项目： | 国家自然科学基金，黑龙江重大攻关课题 |
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| 摘 要： | 通过研究不同生长状态、处理溶液、保存时间及热激时间对感受态细胞制备的影响,分析了影响大肠杆菌株JM109和DH5α感受态细胞形成因素,优化了制备感受态细胞的方法.结果表明,OD600为0.43的大肠杆菌菌液经处理液(20 mmol/L MgCl2 + 80 mmol/L CaCl2)处理,-85℃放置12～14 h,42℃热激处理90 s,转化效率最高,可达5.5×105～6×105 cfu/μg DNA(pSilencer2.1-U6-neo),随着质粒放置时间增加,转化效率下降.
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| 关 键 词： | 感受态细胞 制备方法 转化效率 |
Study on competent cell preparation for increasing efficiency of transformation |
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| WANG Shi-wei,LI Xuye,ZHANG Wei-wei.Study on competent cell preparation for increasing efficiency of transformation[J].Journal of Qiqihar University(Natural Science Edition),2009,25(2). |
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| Authors: | WANG Shi-wei LI Xuye ZHANG Wei-wei |
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| Institution: | 1.Life Science and Engineering College;Qiqihar University;Heilongjiang Qiqihar 161006;China;2.Heilongjiang Animal Husbandry Research Institut;Heilongjiang Qiqihar 161005;China |
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| Abstract: | The affecting factors of forming Escherichia coli competent cells were analyzed to increase transformation efficiency in this paper.A series of competent cells of Escherichia coli were produced in different conditions,including the growth status,the different solutions,the different storing time,hot shock time,plasmid length.And different effects on transformation ability of competent Escherichia coli were evaluated.The result was as follows: The highest transformation ability was obtained when the cells wa... |
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| Keywords: | competent cell preparation optimized protocol transformation efficiency |
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