RESUMO
This overview guides both novices and experienced researchers facing challenging targets to select the most appropriate gene expression system for producing a particular protein. By answering four key questions, readers can determine the most suitable gene expression system following a decision scheme. This guide addresses the most commonly used and accessible systems and provides brief descriptions of the main gene expression systems' key characteristics to assist decision making. Additionally, information has been included for selected less frequently used "exotic" gene expression systems.
Assuntos
Bases de Dados de Produtos Farmacêuticos , Ligantes , Proteínas Recombinantes/genética , Expressão Gênica/genéticaRESUMO
Minimal plasmids play an essential role in many intermediate steps in molecular biology. For example, they can be used to assemble building blocks in synthetic biology or be used as intermediate cloning plasmids that are ideal for PCR-based mutagenesis methods. A small backbone also opens up for additional unique restriction enzyme cloning sites. Here we describe the generation of pICOz, a 1185-bp fully functional high-copy cloning plasmid with an extended multiple cloning site. We believe that this is the smallest high-copy cloning vector ever described.
Assuntos
Clonagem Molecular/métodos , Escherichia coli/genética , Vetores Genéticos/genética , Plasmídeos/genética , Sequência de Bases , Mutagênese , Reação em Cadeia da Polimerase/métodos , Recombinação Genética , Deleção de Sequência , Biologia Sintética/métodosRESUMO
The type VI Secretion System (T6SS) is a tool for Gram-negative pathogens to interact with other bacteria as well as with the eukaryotic host cell. While the role of T6SS in interbacterial interactions has drawn much attention in recent years, research into the T6SS as a human virulence factor continues at a slower pace. Nevertheless, T6SS has been shown to interfere with eukaryotic host cell immunity at several levels, ranging from direct attack of the host cell to attenuation of disease, allowing the pathogen to survive longer in the host environment. In this review, we aim to give a comprehensive overview focused on the ways bacteria use their T6SS in the modulation of mammalian host cell immunity. While doing so, we attempt to describe potential new avenues of research, as well as outline the ways in which T6SS could become a therapeutic target allowing to circumvent existing antibiotic resistance. Although much work remains to be done, a better comprehension of the T6SS mechanisms of action will undoubtedly lead to new strategies to counteract T6SS-bearing pathogens.
