Advanced Bacterial Genetics by John N.Abelson, Melvin I.Simon

By John N.Abelson, Melvin I.Simon

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DESHAIES VOLUME 400. Phase II Conjugation Enzymes and Transport Systems Edited by HELMUT SIES AND LESTER PACKER VOLUME 401. Glutathione Transferases and Gamma Glutamyl Transpeptidases Edited by HELMUT SIES AND LESTER PACKER VOLUME 402. Biological Mass Spectrometry Edited by A. L. BURLINGAME VOLUME 403. GTPases Regulating Membrane Targeting and Fusion Edited by WILLIAM E. BALCH, CHANNING J. DER, AND ALAN HALL VOLUME 404. GTPases Regulating Membrane Dynamics Edited by WILLIAM E. BALCH, CHANNING J.

The hybrid Tn10‐araCþ at the duplication join point illustrated in Fig. 1 [4] identification of essential genes in bacteria 25 connects regions of the chromosome that are normally separated by more than 6000 kbp of DNA. , 1981). We cloned the araCþ gene from E. coli into the HinDIII site of Tn10dTc to create Tn10dTc‐araCþ (plasmid pDH10). Strains defective in the araC gene are complemented by Tn10dTc‐araCþ. This provides a simple screen on arabinose‐utilization plates for the presence or absence of the Tn10dTc‐araCþ derivative.

Plan of the Experiment Isolation of Mud Insertions in and Tn10‐araCþ Near Genes of Interest Mud: The random mutagenesis of Salmonella by MudJ transposition is described in Chapter 7. MudJ transposition is selected by selecting for MudJ‐encoded KmR. When the selection for MudJ insertions is done on rich medium, about 4% of the insertions are in auxotrophic genes. , 1980). For some pathways such as the his biosynthetic genes, all the genes are in a single nine‐gene operon; consequently, the chromosomal location of a his::MudJ insertion is known immediately.

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