Summary
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.
Similar content being viewed by others
References
Adams J, Kinney T, Thompson S, Rubin L, Helling RB (1979) Frequency-dependent selection for plasmid-containing cells of Escherichia coli. Genetics 91:627–637
Anderson EH (1946) Growth requirements of virus-resistant mutants of Escherichia coli stain B. Proc Natl Acad Sci USA 32:120–128
Bender RA, Janssen KA, Resnick AD, Blumenberg M, Foor F, Magasanik B (1977) Biochemical parameters of glutamine synthetase from Klebsiella aerogenes. J Bacteriol 129:1001–1009
Blumenberg M, Magasanik B (1979) A study in evolution: the histidine utilization genes of enteric bacteria. J Mol Biol 135:23–37
Blumenberg M, Magasanik B (1981) Physical maps of Klebsiella aerogenes and Salmonella typhimurium hut genes. J Bacteriol 145:664–667
Boylan SA, Bender RA (1984) Genetic and physical maps of Klebsiella aerogenes genes for histidine utilization (hut). Mol Gen Genet 193:99–103
El-Gewely MR, Helling RB, Farmerie W, Barnett WE (1982) Location of a phenylalanine t-RNA gene on the physical map of the Euglena gracilis chlorolast genome. Gene 17:337–339
El-Gewely MR, Helling RB (1980) Preparative separation of DNA-ethidium bromide complexes by zonal density gradient centrifugation. Anal Biochem 102:423–428
Gilbert W, Maxam A (1973) The nucleotide sequence of the lac operator. Proc Nat Acad Sci USA 70:3581–3584
Goldberg RB, Hanau R (1980) Regulation of Klebsiella pneumoniae hut operons by oxygen. J Bacteriol 141:745–750
Goldberg RB, Magasanik B (1975) Gene order of the histidine utilization (hut) operons in Klebsiella aerogenes. J Bacteriol 122:1025–1031
Goldberg RB, Bloom FR, Magasanik B (1976) Regulation of histidase synthesis in intergeneric hybrids of enteric bacteria. J Bacteriol 127:114–119
Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197
Lederberg EM, Cohen SN (1974) Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol 119:1072–1074
Morrison DA (1977) Transformation in Escherichia coli: cryogenic preservation of competent cells. J Bacteriol 132:349–351
Neidhardt FC, Magasanik B (1957) Reversal of the glucose inhibition of histidase biosynthesis in Aerobacter aerogenes. J Bacteriol 75:253–259
Prival MJ, Magasanik B (1971) Resistance to catabolite repression of histidase and proline oxidase during nitrogen-limited growth of Klebsiella aerogenes. J Biol Chem 246:6288–296
Schlesinger S, Scotto P, Magasanik B (1965) Exogenous and endogenous induction of the histidine-degrading enzymes in Aerobacter aerogenes. J Biol Chem 240:4331–4337
Smith GR (1971) Specialized transduction of the Salmonella hut operons by coliphage λ: Deletion analysis of the hut operons employing λphut. Virology 45:208–223
Struhl K, Cameron JR, Davis RW (1976) Functional genetic expression of eukaryotic DNA in Escherichia coli. Proc Nat Acad Sci USA 73:1471–1475
Sutcliffe JG (1978) Complete nucleotide sequence of Escherichia coli plasmid pBR322. Cold Spring Harbor Symp Quant Biol 43:77–90
Author information
Authors and Affiliations
Additional information
Communicated by G. O'Donovan
Rights and permissions
About this article
Cite this article
Boylan, S.A., Eades, L.J., Janssen, K.A. et al. A restriction enzyme cleavage map of the histidine utilization (hut) genes of Klebsiella aerogenes and deletions lacking regions of hut DNA. Molec. Gen. Genet. 193, 92–98 (1984). https://doi.org/10.1007/BF00327420
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00327420