Abstract
A method is described for measuring the proportion of galactose-specific mRNA (gal-mRNA) in the total RNA extracted from pulse-labelled cells of Escherichia coli K12, by DNA–RNA hybridization with DNA prepared from bacteriophage λdg. RNA from wild-type E. coli was compared with RNA from a homogenote carrying the gal operon both in the chromosome and in a substituted sex-factor, and with RNA from a deletion strain that carried the galactose operon only in the exogenote. In each case the cultures were induced with fucose. Under these conditions the amount of gal-mRNA was found to be proportional to the content of galactokinase in the different cultures, and to the gene frequency. The amounts of gal-mRNA in an Oc mutant and an R− mutant were also proportional to the observed contents of galactokinase. In cultures repressed for the enzymes of the galactose operon with thiomethylgalactoside, the content of gal-mRNA was higher than expected from the content of galactokinase. Possible explanations of this finding are discussed.
Full text
PDF







Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- BUTTIN G. M'ECANISMES R'EGULATEURS DANS LA BIOSYNTH'ESE DES ENZYMES DU M'ETABOLISME DU GALACTOSE CHEZ ESCHERICHIA COLI K12. I. LA BIOSYNTH'ESE INDUITE DE LA GALACTOKINASE ET L'INDUCTION SIMULTAN'EE DE LA S'EQUENCE ENZYMATIQUE. J Mol Biol. 1963 Aug;7:164–182. doi: 10.1016/s0022-2836(63)80044-3. [DOI] [PubMed] [Google Scholar]
- BUTTIN G. M'ECANISMES R'EGULATEURS DANS LA BIOSYNTH'ESE DES ENZYMES DU M'ETABOLISME DU GALACTOSE CHEZ ESCHERICHIA COLI K12. II. LE D'ETERMINISME G'EN'ETIQUE DE LA R'EGULATION. J Mol Biol. 1963 Aug;7:183–205. doi: 10.1016/s0022-2836(63)80045-5. [DOI] [PubMed] [Google Scholar]
- Bishop J. O., Irving M. I. A method for measuring the content of a specific messenger ribonucleic acid in Escherichia coli. Biochem J. 1971 Jan;121(1):105–108. doi: 10.1042/bj1210105. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bishop J. O., Robertson F. W. Transcription of bacteriophage T4 deoxyribonucleic acid in vitro. Biochem J. 1969 Nov;115(3):353–361. doi: 10.1042/bj1150353. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Davison J., Frame R., Bishop J. A partial deletion map of the galactose operon in E. coli K12. Genet Res. 1967 Aug;10(1):107–116. doi: 10.1017/s0016672300010806. [DOI] [PubMed] [Google Scholar]
- Frame R., Bishop J. O. The number of sex-factors per chromosome in Escherichia coli. Biochem J. 1971 Jan;121(1):93–103. doi: 10.1042/bj1210093. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gillespie D., Spiegelman S. A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J Mol Biol. 1965 Jul;12(3):829–842. doi: 10.1016/s0022-2836(65)80331-x. [DOI] [PubMed] [Google Scholar]
- Guha A., Tabaczyński M., Szybalski W. Orientation of transcription for the galactose operon as determined by hybridization of gal mRNA with the separated DNA strands of coliphages lambda-dg. J Mol Biol. 1968 Jul 14;35(1):207–213. doi: 10.1016/s0022-2836(68)80048-8. [DOI] [PubMed] [Google Scholar]
- Ito J., Imamoto F. Sequential derepression and repression of the tryptophan operon in E. coli. Nature. 1968 Nov 2;220(5166):441–444. doi: 10.1038/220441a0. [DOI] [PubMed] [Google Scholar]
- Michaelis G., Starlinger P. Sequential appearance of the galactose enzymes in E. coli. Mol Gen Genet. 1967;100(2):210–215. doi: 10.1007/BF00333607. [DOI] [PubMed] [Google Scholar]
- Morse D. E., Baker R. F., Yanofsky C. Translation of the tryptophan messenger RNA of Escherichia coli. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1428–1435. doi: 10.1073/pnas.60.4.1428. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mueller K., Bremer H. Rate of synthesis of messenger ribonucleic acid in Escherichia coli. J Mol Biol. 1968 Dec;38(3):329–353. doi: 10.1016/0022-2836(68)90390-2. [DOI] [PubMed] [Google Scholar]
- Saedler H., Gullon A., Fiethen L., Starlinger P. Negative control of the galactose operon in E. coli. Mol Gen Genet. 1968;102(1):79–88. doi: 10.1007/BF00341872. [DOI] [PubMed] [Google Scholar]
- Saito S., Ozutsumi M., Kurahashi K. Galactose 1-phosphate uridylyltransferase of Escherichia coli. II. Further purification and characterization. J Biol Chem. 1967 May 25;242(10):2362–2368. [PubMed] [Google Scholar]
- Salser W., Janin J., Levinthal C. Measurement of the unstable RNA in exponentially growing cultures of Bacillus subtilis and Escherichia coli. J Mol Biol. 1968 Jan 28;31(2):237–266. doi: 10.1016/0022-2836(68)90442-7. [DOI] [PubMed] [Google Scholar]
- WILSON D. B., HOGNESS D. S. THE ENZYMES OF THE GALACTOSE OPERON IN ESCHERICHIA COLI. I. PURIFICATION AND CHARACTERIZATION OF URIDINE DIPHOSPHOGALACTOSE 4-EPIMERASE. J Biol Chem. 1964 Aug;239:2469–2481. [PubMed] [Google Scholar]
