Genetic Manipulation of Chlamydia trachomatis: Chromosomal Deletions
| Autor: | Kenneth A. Fields, Mostafa Rahnama, Katerina Wolf |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Mutant
Mutagenesis (molecular biology technique) Chlamydiae Chlamydia trachomatis medicine.disease_cause Article 03 medical and health sciences Transformation Genetic Plasmid medicine Humans Allele Gene Alleles 030304 developmental biology Genetics 0303 health sciences biology 030306 microbiology Chlamydia Infections Chromosomes Bacterial biology.organism_classification Transformation (genetics) Mutagenesis Gene Targeting Gene Deletion |
| Zdroj: | Chlamydia trachomatis ISBN: 9781493996933 Methods Mol Biol |
| DOI: | 10.1007/978-1-4939-9694-0_11 |
| Popis: | Progress in understanding molecular mechanisms contributing to chlamydial pathogenesis has been greatly facilitated by recent advances in genetic manipulation of C. trachomatis. Valuable approaches such as random, chemical-induce mutagenesis or targeted, insertion-based gene disruption have led to significant discoveries. We describe herein a technique for generating definitive null strains via complete deletion of chromosomal genes in C. trachomatis. Fluorescence-reported allelic exchange mutagenesis (FRAEM), using the suicide vector pSUmC, enables targeted deletion of desired chromosomal DNA. The protocol provided here describes steps required to produce transformation competent chlamydiae, generate of a specific allelic exchange plasmid construct, carry out mutagenesis, and isolate clonal populations of resulting mutant strains. |
| Databáze: | OpenAIRE |
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