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Molecular cloning and expression of Neisseria meningitidis class 1 outer-membrane protein in Escherichia coli K-12

Molecular cloning and expression of Neisseria meningitidis class 1 outer-membrane protein in Escherichia coli K-12
Molecular cloning and expression of Neisseria meningitidis class 1 outer-membrane protein in Escherichia coli K-12
A genomic library of meningococcal DNA from a clinical isolate of Neisseria meningitidis was constructed in the expression vector lambda gt11. Outer membrane complex was prepared from the same strain and used to immunize rabbits to raise polyclonal anti-outer membrane complex serum. The amplified library was probed with this polyclonal serum, and seven expressing recombinants were isolated; further investigations indicated these to be identical. The expressed meningococcal gene in these recombinants was fused to vector beta-galactosidase and shown to encode epitopes present on the 42-kilodalton class 1 outer membrane protein. Estimation of the size of the recombinant fusion protein suggests that up to 40 kilodaltons of protein-coding sequence is present. The lambda gt11 recombinant contains a 3.4-kilobase DNA insert, which has been recloned into a plasmid and characterized by restriction endonuclease analysis. A restriction fragment from the insert, representing the protein-coding region hybridizes to a single 2.2-kilobase XbaI fragment from the homologous strain and to similar-sized XbaI fragments in other strains of meningococci, expressing antigenically distinct class 1 proteins
0022-2615
2734-2740
Barlow, A.K.
37084842-5493-4cf1-8405-1d452aa51336
Heckels, J.E.
4f0ffafa-5d84-4fc5-8895-c359d46de0c8
Clarke, I.N.
ff6c9324-3547-4039-bb2c-10c0b3327a8b
Barlow, A.K.
37084842-5493-4cf1-8405-1d452aa51336
Heckels, J.E.
4f0ffafa-5d84-4fc5-8895-c359d46de0c8
Clarke, I.N.
ff6c9324-3547-4039-bb2c-10c0b3327a8b

Barlow, A.K., Heckels, J.E. and Clarke, I.N. (1988) Molecular cloning and expression of Neisseria meningitidis class 1 outer-membrane protein in Escherichia coli K-12. Journal of Medical Microbiology, 55 (11), 2734-2740. (PMID:3117690)

Record type: Article

Abstract

A genomic library of meningococcal DNA from a clinical isolate of Neisseria meningitidis was constructed in the expression vector lambda gt11. Outer membrane complex was prepared from the same strain and used to immunize rabbits to raise polyclonal anti-outer membrane complex serum. The amplified library was probed with this polyclonal serum, and seven expressing recombinants were isolated; further investigations indicated these to be identical. The expressed meningococcal gene in these recombinants was fused to vector beta-galactosidase and shown to encode epitopes present on the 42-kilodalton class 1 outer membrane protein. Estimation of the size of the recombinant fusion protein suggests that up to 40 kilodaltons of protein-coding sequence is present. The lambda gt11 recombinant contains a 3.4-kilobase DNA insert, which has been recloned into a plasmid and characterized by restriction endonuclease analysis. A restriction fragment from the insert, representing the protein-coding region hybridizes to a single 2.2-kilobase XbaI fragment from the homologous strain and to similar-sized XbaI fragments in other strains of meningococci, expressing antigenically distinct class 1 proteins

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Published date: July 1988
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 342330
URI: https://eprints.soton.ac.uk/id/eprint/342330
ISSN: 0022-2615
PURE UUID: 6334eb54-ca6a-4433-baeb-d340b5444c90
ORCID for I.N. Clarke: ORCID iD orcid.org/0000-0002-4938-1620

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Date deposited: 06 Sep 2012 10:43
Last modified: 06 Jun 2018 13:19

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