CONTRUCTION OF DEFINED GALE MUTANTS OF SALMONELLA FOR USE AS VACCINES
CONTRUCTION OF DEFINED GALE MUTANTS OF SALMONELLA FOR USE AS VACCINES
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DOI：
10.1093/infdis/156.1.167
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发表时间：
1987-07-01
影响因子：
6.4
通讯作者：
HACKETT, J
中科院分区：
文献类型：
--
作者：
HONE, D;MORONA, R;HACKETT, J
我们描述了鼠伤寒沙门氏菌LT2半乳糖操纵子的分子克隆和半乳糖启动子及其基因Gale、Galt和Galk的定位。基因序列和转录方向与大肠杆菌K12相同。获得了操纵子的限制性内切酶图谱，并通过转座子插入突变和微细胞分析定位了GAL基因的大致末端。我们构建了一个含有Gale基因0.4kb缺失的质粒，但仍然表达了Gal启动子的GalT和Galk活性。这个Gale缺失被重组到鼠伤寒沙门氏菌和伤寒沙门氏菌TY2的染色体Gal操纵子中。由此产生的菌株是活力旺盛的不可逆转的大风突变株，对半乳糖诱导的0.2 mM半乳糖裂解敏感。鼠伤寒沙门氏菌的衍生物对小鼠无毒，对小鼠有保护作用。
We describe the molecular cloning of the gal operon of Salmonella typhimurium LT2 and the localization of the gal promoter and the genes galE, galT, and galK. The order of the genes and the direction of transcription was the same as in Escherichia coli K12. A restriction enzyme map of the operon was obtained, and the approximate termini of the gal genes were located by using transposon insertion mutagenesis and minicell analysis. We constructed a plasmid that contained a defined 0.4-kilobase deletion in the galE gene, but still expressed galT and galK activities from the gal promoter. This galE deletion was recombined into the chromosomal gal operons of S. typhimurium and Salmonella typhi Ty2. The resulting strains were vigorous, nonreverting galE mutants that were sensitive to galactose-induced lysis at 0.2 mM galactose. The S. typhimurium galE derivatives were avirulent and protective in mice.