Boylan S A, Eades L J, Janssen K A, Lomax M I, Bender R A
Mol Gen Genet. 1984;193(1):92-8. doi: 10.1007/BF00327420.
The histidine utilization (hut) operons of Klebsiella aerogenes were cloned into pBR322. The hut genes are wholly contained on a 7.9 kilobase pair fragment bounded by HindIII restriction sites and expression of hut is independent of the orientation of the fragment with respect to pBR322. A restriction map locating the 27 cleavage sites within hut for the enzymes, HindIII, PvuII, SalI, BglII, KpnI, PstI, SmaI, AvaI, and BamHI was deduced. Several of the cleavage sites for the enzymes HaeIII and HinfI were also mapped. A set of deletion plasmids was isolated by removing various restriction fragments from the original plasmid. These deletions were characterized and were used to assist in mapping restriction sites. This physical characterization of hut DNA opens the way for genetic and molecular analysis of the regulation of hut gene expression in vitro as well as in vivo.
产气克雷伯菌的组氨酸利用(hut)操纵子被克隆到pBR322中。hut基因完全包含在一个由HindIII限制性酶切位点界定的7.9千碱基对片段上,并且hut的表达与该片段相对于pBR322的方向无关。推导了一张限制性酶切图谱,该图谱定位了hut内针对HindIII、PvuII、SalI、BglII、KpnI、PstI、SmaI、AvaI和BamHI这些酶的27个切割位点。还绘制了HaeIII和HinfI这两种酶的几个切割位点图谱。通过从原始质粒中去除各种限制性片段,分离出了一组缺失质粒。对这些缺失进行了表征,并用于辅助绘制限制性酶切位点图谱。hut DNA的这种物理表征为体外和体内hut基因表达调控的遗传和分子分析开辟了道路。