lacⅠ 靶基因突变子的一种正向选择系统的建立  

Development of a selective system for lacⅠ gene mutants

作　　者：黎怀星[1] 卢洋[1] 姚真真[1] 傅继梁[1] 

机构地区：[1]第二军医大学基础医学部医学遗传学教研室,上海200433

出　　处：《第二军医大学学报》1999年第10期713-715,共3页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金;军队"九五"重点课题

摘　　要：目的：建立一种能正向选择ｌａｃⅠ靶基因突变子的选择系统。方法：比较了分别携带有ｌａｃⅠ－ ／ｌａｃＺα型质粒和ｌａｃⅠ＋ ／ｌａｃＺα型质粒的ＤＨ１０Ｂ菌在ＬＢ完全培养基、以葡萄糖为碳源的Ｍ９ 基本培养基和以乳糖为碳源的Ｍ９／Ｌ基本培养基内的生长情况。结果：组成型ＤＨ１０Ｂ菌在Ｍ９／Ｌ固体培养基中只需培养２３ ｈ 便可观察到生长，而诱导型ＤＨ１０Ｂ菌则需培养８８ ｈ 才可观察到生长，但在ＬＢ和Ｍ９ 培养基内两者的生长速度却相同；当将少量的组成型ＤＨ１０Ｂ菌与大量的诱导型ＤＨ１０Ｂ菌混合接种于Ｍ９／Ｌ固体培养基上培养时， Ｍ９／Ｌ固体培养基能从约２．７×１０６ 的ｌａｃⅠ＋ 型ＤＨ１０Ｂ菌中将少量的ｌａｃⅠ－ 型ＤＨ１０Ｂ菌筛选出来。结论：本研究建立了一种在特定时间内只选择生长携带有ｌａｃⅠ－ ／ｌａｃＺα型质粒的ＤＨ１０Ｂ菌落。Objective:To develop a selective system for positive selection of lacⅠ mutants. Methods: Investigations on the growth of DH10B cells harboring lacⅠ - or lacⅠ + genes in LB, M9 and M9/L culture media were carried out in this study. Results: After 23 h of culture DH10B cells harboring lacⅠ - genes could grow and form colonies in the M9/L solid medium, and 88 h culture was essential for the DH10B cells harboring lacⅠ + gene to grow into colonies in the M9/L solid media. However, no obvious difference was observed on the growth of DH10B cells harboring lacⅠ - genes or lacⅠ + genes. The results also showed that isolated DH10B cells harboring lacⅠ - genes could be screened from 2.7×10 6 of DH10B cells with lacⅠ + genes based on the M9/L solid media. Conclusion:A selective system, based on the M9/L medium and DH10B host cells, for positive selection of lacⅠ mutants is established in our study.

关 键 词：正向选择 lacⅠ靶基因 基因突变子 

分 类 号：Q754[生物学—分子生物学]