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.

Corynebacterium glutamicum was discovered in Japan in 1956 as a natural glutamate producer. Its “microbial factory” qualities, such as its physiological plasticity and robust catalytic functionalities, have since facilitated the development of efficient production processes for amino acids, nucleotides and vitamins. This monograph illustrates how the information gleaned from complete genome sequencing allows the rational engineering of the entire cellular metabolism and how systems biology permits the further optimization of C. glutamicum as a biocatalyst. Aspects of gene regulation, metabolic pathways, sugar uptake, protein secretion, cell division and biorefinery applications highlight the enormous biotechnological and biorefinery potential.

Diagnostic Molecular Biology, Second Edition describes the fundamentals of molecular biology in a clear, concise manner with each technique explained within its conceptual framework and current applications of clinical laboratory techniques comprehensively covered. This targeted approach covers the principles of molecular biology, including basic knowledge of nucleic acids, proteins and chromosomes; the basic techniques and instrumentations commonly used in the field of molecular biology, including detailed procedures and explanations; and the applications of the principles and techniques currently employed in the clinical laboratory. Topics such as whole exome sequencing, whole genome sequencing, RNA-seq, and ChIP-seq round out the discussion. Fully updated, this new edition adds recent advances in the detection of respiratory virus infections in humans, like influenza, RSV, hAdV, hRV but also corona. This book expands the discussion on NGS application and its role in future precision medicine. - Provides explanations on how techniques are used to diagnosis at the molecular level - Explains how to use information technology to communicate and assess results in the lab - Enhances our understanding of fundamental molecular biology and places techniques in context - Places protocols into context with practical applications - Includes extra chapters on respiratory viruses (Corona)

This book describes the structures and functions of active protein filaments, found in bacteria and archaea, and now known to perform crucial roles in cell division and intra-cellular motility, as well as being essential for controlling cell shape and growth. These roles are possible because the cytoskeletal and cytomotive filaments provide long range order from small subunits. Studies of these filaments are therefore of central importance to understanding prokaryotic cell biology. The wide variation in subunit and polymer structure and its relationship with the range of functions also provide important insights into cell evolution, including the emergence of eukaryotic cells. Individual chapters, written by leading researchers, review the great advances made in the past 20-25 years, and still ongoing, to discover the architectures, dynamics and roles of filaments found in relevant model organisms. Others describe one of the families of dynamic filaments found in many species. The most common types of filament are deeply related to eukaryotic cytoskeletal proteins, notably actin and tubulin that polymerise and depolymerise under the control of nucleotide hydrolysis. Related systems are found to perform a variety of roles, depending on the organisms. Surprisingly, prokaryotes all lack the molecular motors associated with eukaryotic F-actin and microtubules. Archaea, but not bacteria, also have active filaments related to the eukaryotic ESCRT system. Non-dynamic fibres, including intermediate filament-like structures, are known to occur in some bacteria.. Details of known filament structures are discussed and related to what has been established about their molecular mechanisms, including current controversies. The final chapter covers the use of some of these dynamic filaments in Systems Biology research. The level of information in all chapters is suitable both for active researchers and for advanced students in courses involving bacterial or archaeal physiology, molecular microbiology, structural cell biology, molecular motility or evolution. Chapter 3 of this book is open access under a CC BY 4.0 license.

Lasso peptides form a growing family of fascinating ribosomally-synthesized and post-translationally modified peptides produced by bacteria. They contain 15 to 24 residues and share a unique interlocked topology that involves an N-terminal 7 to 9-residue macrolactam ring where the C-terminal tail is threaded and irreversibly trapped. The ring results from the condensation of the N-terminal amino group with a side-chain carboxylate of a glutamate at position 8 or 9, or an aspartate at position 7, 8 or 9. The trapping of the tail involves bulky amino acids located in the tail below and above the ring and/or disulfide bridges connecting the ring and the tail. Lasso peptides are subdivided into three subtypes depending on the absence (class II) or presence of one (class III) or two (class I) disulfide bridges. The lasso topology results in highly compact structures that give to lasso peptides an extraordinary stability towards both protease degradation and denaturing conditions. Lasso peptides are generally receptor antagonists, enzyme inhibitors and/or antibacterial or antiviral (anti-HIV) agents. The lasso scaffold and the associated biological activities shown by lasso peptides on different key targets make them promising molecules with high therapeutic potential. Their application in drug design has been exemplified by the development of an integrin antagonist based on a lasso peptide scaffold. The biosynthesis machinery of lasso peptides is therefore of high biotechnological interest, especially since such highly compact and stable structures have to date revealed inaccessible by peptide synthesis. Lasso peptides are produced from a linear precursor LasA, which undergoes a maturation process involving several steps, in particular cleavage of the leader peptide and cyclization. The post-translational modifications are ensured by a dedicated enzymatic machinery, which is composed of an ATP-dependent cysteine protease (LasB) and a lactam synthetase (LasC) that form an enzymatic complex called lasso synthetase. Microcin J25, produced by Escherichia coli AY25, is the archetype of lasso peptides and the most extensively studied. To date only around forty lasso peptides have been isolated, but genome mining approaches have revealed that they are widely distributed among Proteobacteria and Actinobacteria, particularly in Streptomyces, making available a rich resource of novel lasso peptides and enzyme machineries towards lasso topologies.