Bacteriophage MB78 DNA synthesis is specifically inhibited by the chelating agent ethylene diamine tetraacetic acid

Verma, Mukesh; Chakravorty, Maharani

doi:10.1111/j.1574-6968.1987.tb02253.x

Cited by 2 publications

(1 citation statement)

References 14 publications

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“…It cannot multiply in minimal medium containing citrate. The chelating agent EDTA is an effective inhibitor of its DNA synthesis, whereas EGTA and orthophenanthroline have practically no effect on the development of the phage (4,5). It is a dominant phage, in the sense that it does not allow other phages, such as P22 and 9NA, to grow in its presence.…”

Section: Introductionmentioning

confidence: 99%

Molecular Cloning, Sequencing, and Expression of Two Late Proteins of Bacteriophage MB78

Kolla¹,

Datta²,

Chakravorty³

1999

IUBMB: Life

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Bacteriophage MB78, a virulent phage of Salmonella typhimurium, does not allow other phages, such as P22 and 9NA, to grow in its presence. A detailed physical map of this phage has been constructed in our laboratory. In an ongoing effort to understand the genetics of this interesting phage, various genes were characterized. Here, we report cloning, sequencing, and expression of two late proteins, coded in a SalI-HindIII fragment (SH9), by using the minicell expression system. Further, we performed a kinetic study of phage proteins by infection the host LT2 cells and compared the proteins produced, with proteins obtained by the minicell expression system. Both sets of proteins run exactly parallel and migrated as 14- and 15-kDa proteins on a polyacrylamide gel. The synthesis of these two proteins started 15 min after infection with MB78 and was prominent after 45 min. One of the proteins exhibited 57% homology to the structural protein of mycobacteriophage L5.

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Section: Introductionmentioning

confidence: 99%

Molecular Cloning, Sequencing, and Expression of Two Late Proteins of Bacteriophage MB78

Kolla¹,

Datta²,

Chakravorty³

1999

IUBMB: Life

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Synthesis of a bacteriophage MB78 late protein by novel ribosomal frameshifting

Kolla

Chakravorty

Pandey

et al. 2000

Gene

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MB78 is a virulent phage of Salmonella typhimurium that possesses a number of interesting features, making it a suitable organism to study the regulation of gene expression. A detailed physical map of this phage genome has been constructed and is being extensively studied at the molecular level. Here, we demonstrate the expression of two late proteins of bacteriophage MB78 derived from the same gene as a result of possible ribosomal frameshifting. In vitro transcription-translation yields a major protein that migrates as 28kDa, whereas in vivo expression using pET expression vectors yields two equally expressed proteins of molecular sizes 28 and 26kDa. A putative slippery sequence TTTAAAG and a pseudoknot structure, two essential cis elements required for the classical ribosomal frameshifting, are identified in the reading frame. Mutations created at the slippery sequence resulted in a single 28kDa protein and completely abolished the expression of 26kDa protein. Thus, we have produced the first evidence that ribosomal frameshifting occurs in bacteriophage MB78 of Salmonella typhimurium.

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Bacteriophage MB78 DNA synthesis is specifically inhibited by the chelating agent ethylene diamine tetraacetic acid

Cited by 2 publications

References 14 publications

Molecular Cloning, Sequencing, and Expression of Two Late Proteins of Bacteriophage MB78

Molecular Cloning, Sequencing, and Expression of Two Late Proteins of Bacteriophage MB78

Synthesis of a bacteriophage MB78 late protein by novel ribosomal frameshifting

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