Construction of New Campylobacter Cloning Vectors and a New Mutational Cat Cassette
Abstract
We have developed new Campylobacter shuttle vectors which are 6.5-6. 8-kb plasmids carrying Campylobacter and Escherichia coli replicons, a multiple cloning site (MCS), the lacZ(alpha) gene, oriT and either a kanamycin or chloramphenicol resistance-encoding gene (Km(R) or Cm(R)) from Campylobacter which functions in both hosts. These vectors can be mobilized efficiently from E. coli into C. jejuni or C. coli, and stably maintained in these hosts. Plasmids pRY107 and pRY108 carry a Km(R) marker and 17 unique cloning sites in two different orientations in lacZ(alpha), allowing easy blue/white color selection. Plasmids pRY111 and pRY112 contain a Cm(R) gene and 17 unique sites in both orientations. In addition, MCS are flanked by T7 and T3 late promoters and M13 forward and reverse primer sites, facilitating expression in T7 or T3 expression systems and sequence analysis. A Campylobacter Cm(R) gene cartridge, bracketed by six restriction sites, has been developed for use in site-specific mutagenesis of Campylobacter genes.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 1993
- Accession Number
- ADA275605
Entities
People
- Patricia Guerry
- Richard A. Alm
- Ruijin Yao
- Trevoir J. Trust
Organizations
- Naval Medical Research Center