Widera G, Gautier F, Lindenmaier W, Collins J
Mol Gen Genet. 1978 Jul 25;163(3):301-5. doi: 10.1007/BF00271959.
In vitro recombination techniques were used to construct hybrid plasmids between pBR 322 and different DNA fragments derived from pML 21 and the E. coli chromosome. Some of the resulting hybrid plasmids express the tetracycline resistance gene of pBR 322, depending on the DNA fragment which has been ligated into the HindIII-site of pBR 322. From these studies we could conclude the direction of transcription of the kanamycin resistance gene in plasmid pML 21 with respect to the SalI, SmaI, HincII, KpnI, EcoRI and HindIII restriction sites. The replacement of the small HindIII-EcoRI-fragment in pBR 322 by other DNA fragments from the E. coli chromosome and selection for tcr-phenotypes showed that this system may be very useful for screening and analysis of promotor-containing DNA fragments.
利用体外重组技术构建了pBR 322与来源于pML 21和大肠杆菌染色体的不同DNA片段之间的杂种质粒。根据连接到pBR 322的HindIII位点的DNA片段，一些所得的杂种质粒表达pBR 322的四环素抗性基因。通过这些研究，我们可以确定质粒pML 21中卡那霉素抗性基因相对于SalI、SmaI、HincII、KpnI、EcoRI和HindIII限制性位点的转录方向。用大肠杆菌染色体的其他DNA片段取代pBR 322中的小HindIII-EcoRI片段并选择四环素抗性表型，表明该系统对于含启动子的DNA片段的筛选和分析可能非常有用。
