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M S Malo

Publications and source records attributed to M S Malo.

5 recordsLinked to original sources
PubMed1990

Promoter elements and regulation of expression of the cysD gene of Escherichia coli K-12.

The cysD gene, involved in cysteine biosynthesis in Escherichia coli and Salmonella typhimurium, is positively regulated by the CysB regulatory protein. The cysD promoter of E. coli K-12 in a 492-bp PstI-Eco RI fragment was sequenced. The in vivo transcription start point (tsp) for the cysD gene was determined by the methods of T4 DNA polymerase mapping and mung-bean nuclease mapping. The -10 region of the cysD promoter (TATAGT) is closely homologous to the -10 consensus sequence (TATAAT) for E. coli promoters. The -35 region of this promoter (TTCATT) is less closely related to the -35 consensus sequence (TTGACA). Several mutants were obtained by using a chain-termination method for generating unidirectional deletions. Evidence is presented for a possible CysB protein binding site around -89, thought to be involved in regulation of expression of the cysD gene.

Amino Acid Sequence↗
PubMed1988

Promoter-detection vectors for Escherichia coli with multiple useful features.

Two promoter-detection vectors have been constructed which enable the cloning and characterization of promoters recognized by the RNA polymerase of Escherichia coli K-12. The intergenic region of phage M13 DNA, present in opposite orientations in the two vectors, permits the preparation of single-stranded DNA of either strand of the insert thus facilitating oligodeoxyribonucleotide heteroduplex mutagenesis and sequencing of both strands by the dideoxy method of chain termination. After mutagenesis, isolates can be screened for changed function by replica-plating colonies to plates containing XGal. Selected isolates can be characterized by nucleotide sequence analysis, to determine the change in structure, and by beta-galactosidase assays, thus measuring the effect of mutagenesis on promoter function. The vectors could also be used like other protein-fusion vectors.

Base Sequence↗
PubMed1987

Lambda transducing phage and clones carrying genes of the cysJIHDC gene cluster of Escherichia coli K12.

DNA from each of two specialized transducing lambda phage, lambda dcysJIHD and lambda cysJ, has been analysed by heteroduplex mapping. The segment of the Escherichia coli chromosome carried by lambda dcysJIHD was shown to be large, approximately 18 kb in length, and to replace a large length of lambda DNA, approximately 11 kb, which includes the genes for integration and recombination. Thus lambda dcysJIHD is a bio-type transducing phage. lambda cysJ was shown to have lost very little lambda DNA and to carry about 8 kb of bacterial DNA. Sites for several restriction endonucleases were mapped in the DNA from each phage and cloning experiments located some of the genes of the cluster in relation to the restriction map. Cysteine regulation of the cloned cysJ and cysD genes was shown as well as cysteine regulation of beta-galactosidase in some constructs. The direction of transcription of the cysD gene was established, and from physical evidence the size of the 'silent section' between the cysH and cysD genes was estimated to be at least 11 kb.

Bacteriophage lambda↗