酵母单杂交诱饵质粒pHis-3NF和pLacZ-3NF的构建与鉴定

doi:10.3969/j.issn.1004-616x.2003.01.004

癌变·畸变·突变 ›› 2003, Vol. 15 ›› Issue (1): 13-16.doi: 10.3969/j.issn.1004-616x.2003.01.004

酵母单杂交诱饵质粒pHis-3NF和pLacZ-3NF的构建与鉴定

牛永东1, 许丽艳2, 李恩民1

1. 汕头大学医学院生物化学与分子生物学教研室；　2. 汕头大学医学院肿瘤病理研究室，广东　汕头　515031

收稿日期:2002-10-31 修回日期:2002-11-26 出版日期:2003-01-30 发布日期:2003-01-30
通讯作者: 李恩民

CONSTRUCTION AND IDENTIFICATION OF THE BAIT PLASMIDS pHis-3NF AND pLacZ-3NF FOR YEAST ONE-HYBRID SYSTEM

NIU Yong-dong 1, XU Li-yan 2, LI En-min 1

1. Department of Biochemistry ＆ Molecular Biology; 2. Insititue of Oncologic Pathology,Medical College of Shantou University, Shantou 515031 , China

Received:2002-10-31 Revised:2002-11-26 Online:2003-01-30 Published:2003-01-30
Contact: LI En-min

摘要/Abstract

摘要： 目的：构建可以与酵母染色体组发生重组的质粒pHis-NF和pLacZ-NF。方法：人工合成44 bp的三重NF-κB元件寡聚核苷酸，通过DNA重组技术将靶序列NF-κB串定向插入细菌-酵母穿梭质粒pHisi和pLacZi的多克隆位点MCS后，用特异性酶切、琼脂糖凝胶电泳、PAGE电泳、DNA测序等方法进行鉴定。结果： PAGE电泳和DNA测序分析证实定向插入的寡聚核苷酸串与预先设计的插入序列一致。结论：成功地构建了重组质粒pHis-3NF和pLacZ-3NF，为借助酵母单杂交实验技术平台深入探讨NF-κB因子相关基因的转录调控机制奠定了实验材料和技术基础。

关键词: 核转录因子, NF-κB, pHis-3NF, pLacZ-3NF, 酵母单杂交

Abstract: Purpose: To construct the pHis-NF and pLacZ-NF plasmids which could be integrated into the yeast genome. Methods: the target DNA sequence of tandem NF-κB copies was synthesized, and the tandem copies were directionally inserted into the MCS of E.coli yeast shuttle pHisi and pLacZi plasmid vectors respectively. The recombinant were confirmed by restriction enzyme digestion, agarose gel, PAGE electropheresis and DNA sequencing. Results: Both PAGE electropheresis and DNA sequence analysis confirmed the inserted oligonucleotides. Conclusion: These two plasmids, pHis-3NF and pLacZ-3NF have been successfully constructed.

Key words: Nuclear factors, NF-κB, pHis-3NF, pLacZ-3NF, Yeast one-hybrid system

牛永东, 许丽艳, 李恩民. 酵母单杂交诱饵质粒pHis-3NF和pLacZ-3NF的构建与鉴定[J]. 癌变·畸变·突变, 2003, 15(1): 13-16.

NIU Yong-dong , XU Li-yan , LI En-min . CONSTRUCTION AND IDENTIFICATION OF THE BAIT PLASMIDS pHis-3NF AND pLacZ-3NF FOR YEAST ONE-HYBRID SYSTEM[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2003, 15(1): 13-16.

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酵母单杂交诱饵质粒pHis-3NF和pLacZ-3NF的构建与鉴定

CONSTRUCTION AND IDENTIFICATION OF THE BAIT PLASMIDS pHis-3NF AND pLacZ-3NF FOR YEAST ONE-HYBRID SYSTEM

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