Chen J W, Bennett D C, Umbarger H E
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907.
J Bacteriol. 1991 Apr;173(7):2328-40. doi: 10.1128/jb.173.7.2328-2340.1991.
Three different approaches were used to examine the regulatory effects of the amino acids specified by the peptide-coding region of the leader transcript of the ilvGMEDA operon of Escherichia coli K-12. Gene expression was examined in strains carrying an ilvGMED'-lac operon fusion. In one approach, auxotrophic derivatives were starved of single amino acids for brief periods, and the burst of beta-galactosidase synthesis upon adding the missing amino acid was determined. Auxotrophic derivatives were also grown for brief periods with a limited supply of one amino acid (derepression experiments). Finally, prototrophic strains were grown in minimal medium supplemented with single and multiple supplements of the chosen amino acids. Although codons for arginine, serine, and proline are interspersed among the codons for the three branched-chain (regulatory) amino acids, they appeared to have no effect when added in excess to prototrophs or when supplied in restricted amounts to auxotrophs. Deletions removing the terminator stem from the leader removed all ilv-specific control, indicating that the attenuation mechanism is the sole mechanism for ilv-specific control.
采用三种不同方法研究了大肠杆菌K - 12的ilvGMEDA操纵子前导转录本肽编码区所指定的氨基酸的调控作用。在携带ilvGMED'-lac操纵子融合体的菌株中检测基因表达。在一种方法中,将营养缺陷型衍生物短期饥饿处理单一氨基酸,然后测定添加缺失氨基酸后β - 半乳糖苷酶合成的爆发情况。营养缺陷型衍生物也在一种氨基酸供应有限的情况下短期培养(去阻遏实验)。最后,原养型菌株在补充了所选氨基酸单一或多种添加物的基本培养基中生长。尽管精氨酸、丝氨酸和脯氨酸的密码子散布在三种支链(调控)氨基酸的密码子之间,但当向原养型菌株过量添加这些氨基酸或向营养缺陷型菌株限量供应时,它们似乎没有影响。从前导序列中去除终止子茎的缺失突变消除了所有ilv特异性调控,表明衰减机制是ilv特异性调控的唯一机制。