| 快速LacZ检测法提高酵母双杂交试验的敏感度 |
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| 引用本文： | 郭佳岩,宋婀莉,马素参,高友鹤. 快速LacZ检测法提高酵母双杂交试验的敏感度[J]. 生物技术, 2004, 14(1): 31-33 |
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| 作者姓名： | 郭佳岩 宋婀莉 马素参 高友鹤 |
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| 作者单位： | 中国医学科学院基础医学研究所,中国协和医科大学基础医学院,中国医学科学院蛋白质组学研究中心,北京,100005 |
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| 基金项目： | 国家自然科学基金(30270657);国家高技术研究发展计划(863)(2001AA233051);国际科技合作重点项目计划(2002AA229031);重大基础研究前期研究专项项目计划(2002CCA04100). |
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| 摘 要： | 目的：建立快速简单敏感准确的LacZ检测法。方法：将在SD/-Trp-Leu培养液中过培养的待检测酵母菌直接点样在滤纸上进行LacZ检测，筛选出LacZ阳性克隆。结果：将小鼠3T3文加转化含有WWP1的酵母菌，选出10个较大的SD/-Trp-Leu-His培养阳性的克隆，利用快速LacZ检测法检测这10个酵母克隆，结果在5h内不同程度地变蓝，呈阳性结果；而作为对照的传统检测方法在8h内的检测结果均显示为阴性。结论：采用快速LacZ检测法检测蛋白质之间的相互作用可以大大缩短筛选的时间，简化筛选的步骤，增加检测的敏感度和准确性。
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| 关 键 词： | 传统LaeZ检测法 酵母双杂交技术 蛋白质相互作用 快速LacZ检测法 |
| 文章编号： | 1004-311X(2004)01-0031-03 |
| 修稿时间： | 2003-07-03 |
Rapid LacZ Screening Method Increases the Sensitivity of Yeast Two-hybrid Screening |
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| GUO Jia-yan,SONG E-li,MA Su-can,GAO You-he. Rapid LacZ Screening Method Increases the Sensitivity of Yeast Two-hybrid Screening[J]. Biotechnology, 2004, 14(1): 31-33 |
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| Authors: | GUO Jia-yan SONG E-li MA Su-can GAO You-he |
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| Affiliation: | GUO Jia-yan,SONG E-li,MA Su-can,GAO You-he~* |
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| Abstract: | Objective:To establish rapid,simple,sensitive,and accurate LacZ screening method.Methods:Yeasts grown in SD/-Trp-Leu liquid culture medium were directly put on the filter paper for LacZ screening for positive clones.Result:Mouse 3T3 library was transformed to the yeasts,which contain the protein WWP1,and 10 big clones grown in SD/-Trp-Leu-His agar were identified by two LacZ screening methods.All the clones turned blue (i.e.positive results) in 5 hours by the rapid LacZ method,but none of the 10 clones turned blue in 8 hours by the traditional LacZ method.Conclusion:Rapid LacZ method identifies positive clones with less time and fewer steps than traditional method.And the sensitivity and accuracy can be great improved. |
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| Keywords: | Rapid LacZ method Traditional LacZ method Yeast two-hybrid Proteins interaction |
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