Molecular Cloning and Gene Regulation in Bacilli

1st Edition - January 28, 1982
Editor: A Ganesan
Language: English
eBook ISBN:
9 7 8 - 0 - 3 2 3 - 1 5 3 0 9 - 6

Molecular Cloning and Gene Regulation in Bacilli presents the proceedings of the 1981 Cetus Conference on Genetics held at Stanford University, Stanford, California. It summarizes… Read more

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Molecular Cloning and Gene Regulation in Bacilli presents the proceedings of the 1981 Cetus Conference on Genetics held at Stanford University, Stanford, California. It summarizes both basic and applied aspects of bacilli genetics. It discusses significant advances made in understanding chromosome structure, gene arrangement, molecular cloning, cloned gene expression, DNA metabolism, transcription, and translation. Divided into five sessions, the book starts by discussing the DNA sequence from RNA intergenic spaces of Bacillus subtilis rRNA gene sets, the construction of a bifunctional cosmid vector of large DNA segments, and the mating system of bacilli. Molecular cloning session covers complementation system and dominance analyses in Bacillus, genetic fusion of Escherichia coli lac genes to a Bacillus subtilis promoter, and DNA cloning of B. subtilis. It also describes the construction of trimethoprim resistant B. subtilis plasmid and expression of E. coli trp genes cloned in B. subtilis. Session III encompasses chapters that discuss protein secretion by bacilli; regulation of alpha-amylase production in B. subtilis; entomocidal toxin translation of B. thuringiensis; and expression of crystal protein, heterologous, and eukaryotic genes in bacilli. Session IV focuses on various aspects of DNA metabolism of bacilli, such as the interaction of bacterial chromosome with cell membrane; plasmid DNA in competent cells and protoplasts of B. subtilis; analysis of peptides synthesized by B. subtilis mutants; and DNA repair, uptake, restriction, modification, and recombination. The final session examines species-specific translation, control of gene expression and replications in plasmids, development of expression-vector in B. subtilis, and regulatory modifications of RNA polymerase. Each chapter is presented in an experimental manner, consisting of a summary of the study, materials and methods, results, as well as references.

Contributors

Preface

Opening Remarks

Session I Chromosome Structure and Gene Arrangement

Nucleotide Sequence of Intergenic Spacer DNA of Three 16S-23S RNA Gene Sets

from B. subtilis

Use of a Bifunctional Cosmid for Cloning Large DNA Fragments

of B. subtilis

B. subtilis Mating System: Dicaryotic Recombinants and Suppression of the

Prototrophic Phenotype

Genetic Manipulations with Phage SPB

Session II Molecular Cloning

System for Complementation and Dominance Analyses in Bacillus

Genetic Fusion of E. coli lac Genes to a B. subtilis Promoter

DNA Cloning in B. subtilis. IV. Transformation of Competent Cells by Internally

Repeated Plasmids

Construction of a Trimethoprim Resistant B. subtilis Plasmid for Use in Cloning the E. coli trpC Gene in B. subtilis

Expression of E. coli trp Genes Cloned in B. subtilis

Session III Expression of Cloned Genes

Penicillinase and the Secretion of Proteins by Bacilli

Regulation of α-Amylase Production in B. subtilis

In Vitro Translation of the Entomocidal Toxin of B. thuringiensis

Cloning the Crystal Protein Gene of B. thuringiensis in E. coli

Expression and Instability of Heterologous Genes in B. subtilis

Expression of Eukaryotic Genes in B. subtilis Using Signals of penP

Session IV DNA Metabolism

Initiation of Chromosome Replication and Membrane Association of the Replication Origin in B. subtilis

Characterization of the Chromosomal Terminus of B. subtilis and its Attachment to the Cell Membrane

Characterization of the Terminus Region of the B. subtilis Chromosome

Fate of Plasmid DNA in Competent Cells and Protoplasts of B. subtilis

Coordinated Changes in Competence Associated Polypeptides among Transformation Defective Mutants of B. subtilis

DNA Repair in B. subtilis: The Identification of a Strain Temperature Inducible for 'SOS' Functions

DNA-Dependent ATPases in B. subtilis: Properties and Functions

Uptake, Restriction, Modification and Recombination of DNA Molecules During Transformation in B. subtilis

Session V. Transcription and Translation

Species Specific Translation: Characterization of B. subtilis Ribosome Binding Sites

Control of Gene Expression and Replication in Plasmids pE194 and pC194

The Development of an Expression-Vector in B. subtilis Using a Heterologous Gene

Anatomy of a Sporulation Gene: Nucleotide Sequences that Signal the Initiation of Transcription and Translation

Biochemical Approaches to the Study of Regulatory Modifications of RNA Polymerase; Identification and Characterization of B. subtilis Sigma-28 RNA Polymerase, an Enzyme of Unique Promoter Specificity

The Transition between Early and Middle Gene Expression in the Development of Phage SP01: Physiological and Biochemical Aspects