Localization of the structural gene of colicin A on the restriction map of the plasmid pCol A—CA 31 through hybridization with its messenger RNA

Morlon, Juliette; Cavard, D; Lazdunski, Claude

doi:10.1016/0014-5793(82)80028-8

Cited by 5 publications

(1 citation statement)

References 8 publications

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“…We had previously shown that the gene caa is entirely included in the 2.17-kb HincII fragment of plasmid ColA (Morlon et al, 1982b). Two PstI sites are located in this fragment (Morlon et al, 1982a).…”

Section: Resultsmentioning

confidence: 99%

Nucleotide sequence of promoter, operator and amino-terminal region of caa, the structural gene of colicin A.

Morlon¹,

Lloubès²,

Chartier³

et al. 1983

The EMBO Journal

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Section: Resultsmentioning

confidence: 99%

Nucleotide sequence of promoter, operator and amino-terminal region of caa, the structural gene of colicin A.

Morlon¹,

Lloubès²,

Chartier³

et al. 1983

The EMBO Journal

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Lysis protein encoded by plasmid ColA-CA31

et al. 1985

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A gene, cal, coding for a polypeptide needed for the release of colicin A from Escherichia coli cells has been identified by transposon insertion. The cal gene was located on the ColA plasmid map adjacent to cai, the gene coding for colicin A immunity protein, and therefore 592 bases downstream from caa, the structural gene for colicin A. Transcription of cal is in the same direction as caa, that is in the opposite direction to cai. Its sequence has been determined and the predicted amino acid composition features a basic N-terminal end followed by a serie of hydrophobic residues similar to the signal sequence in precursors of exported proteins. The C-terminal part also contains a core of hydrophobic residues. The overall amino acid sequence of the cal protein is homologous to that of lytic proteins encoded by the related plasmids pColE1, and pCloDF13. The cal protein has been identified on urea-SDS-polyacrylamide gels by selective labelling with various radioactive amino acids and its synthesis is co-induced with that of colicin A. The cal protein undergoes slow processing with loss of the N-terminal "signal" region and the mature form is released into the medium together with colicin A.

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Localization of genes responsible for replication and immunity to colicin A on plasmid ColA-CA31

1983

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The region containing the origin and regulatory sites for replication as well as the immunity gene (iaa) have been localized on the plasmid ColA-CA31. The region involved in replication functions of ColA can be hybridized with that of ColE1. It is located between 1 and 1 kb on the plasmid map previously published (Morlon et al. 1982a). A 0.50 Kb HincII fragment of ColA can be weakly hybridized to the ColE1 immunity region. This fragment contains iaa since directed in vitro mutagenesis at an internal restriction site can abolish the immunity to colicin A; however, it does not contain the entire iaa. Knowing the localization of regions involved in autonomous DNA replication and immunity, a mini-ColA plasmid was constructed that contains these two regions. The mini-ColA of 2.8 Kb can be amplified in the presence of chloramphenicol and confers the immunity to transformants. It thus constitutes a useful cloning vector. Expression of ColA and of the various constructed plasmids in the maxicell system suggests that the immunity protein has a molecular weight of about 18-20 Kd.

show abstract

Localization of the structural gene of colicin A on the restriction map of the plasmid pCol A—CA 31 through hybridization with its messenger RNA

Cited by 5 publications

References 8 publications

Nucleotide sequence of promoter, operator and amino-terminal region of caa, the structural gene of colicin A.

Nucleotide sequence of promoter, operator and amino-terminal region of caa, the structural gene of colicin A.

Lysis protein encoded by plasmid ColA-CA31

Localization of genes responsible for replication and immunity to colicin A on plasmid ColA-CA31

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