Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1986 Jan;165(1):21-7.
doi: 10.1128/jb.165.1.21-27.1986.

Cloning, nucleotide sequencing, and expression of tetanus toxin fragment C in Escherichia coli

Cloning, nucleotide sequencing, and expression of tetanus toxin fragment C in Escherichia coli

N F Fairweather et al. J Bacteriol. 1986 Jan.

Abstract

The amino acid sequence of the first 30 residues of fragment C of tetanus toxin was determined, and a mixture of 32 complementary oligonucleotides, each 17 bases long, was synthesized. A 2-kilobase (kb) EcoI fragment of Clostridium tetani DNA was identified by Southern blotting and was cloned into the Escherichia coli plasmid vector pAT153 with the 32P-labeled oligonucleotide mixture as a probe. A second 3.2-kb Bg/II fragment was identified and cloned with the 2-kb EcoRI fragment as a probe. The nucleotide sequence of 1.8 kb of this DNA was determined and was shown to encode the entire fragment C and a portion of fragment B of tetanus toxin. The tetanus DNA was expressed in E. coli with pWRL507, a plasmid vector containing the trp promoter and a portion of the trpE gene. The trpE-tetanus fusion proteins were visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and were shown to react with anti-fragment C antibody.

PubMed Disclaimer

References

    1. Infect Immun. 1975 Nov;12(5):1147-53 - PubMed
    1. J Mol Biol. 1975 Nov 5;98(3):503-17 - PubMed
    1. FEBS Lett. 1976 Sep 15;68(1):5-7 - PubMed
    1. J Biol Chem. 1977 Jan 10;252(1):187-93 - PubMed
    1. J Biol Chem. 1977 Jan 10;252(1):194-8 - PubMed

MeSH terms

Associated data

LinkOut - more resources