Gene Transfer and Expression of Recombinant Proteins in Moderately Halophilic Bacteria
Moderately halophilic bacteria (MHB) of the genera Halomonas and Chromohalobacter have been used as hosts for the expression of heterologous proteins of biotechnological interest, thus expanding their potential to be used as cell factories for various applications. This chapter deals with the methodology for the construction of recombinant plasmids, their transfer to a number of MHB, and the assaying of the corresponding heterologous proteins activity. The transferred genes include (1) inaZ , encoding the ice nucleation protein of the plant pathogen Pseudomonas syringae , (2) gfp , encoding a green fluorescent protein from the marine bioluminescent jellyfish Aequorea victoria , and (3) the α -amylase gene from the hyperthermophilic archeon Pyrococcus woesei . Vector pHS15, which was designed for expression of heterologous proteins in both E. coli and MHB, was used for the subcloning and transfer of the above genes. The recombinant constructs were introduced to MHB by assisted conjugal transfer from E. coli donors. The expression and function of the recombinant proteins in the MHB transconjugants is described.
- Dendrimer-Mediated Cell Transfection in Vitro
- Generating Conditional Knockout Mice
- Recombinant Protein Production in Plants: Challenges and Solutions
- Cloning into M13
- DNA Double-Strand Break Damage and Repair Assessed by Pulsed-Field Gel Electrophoresis
- In Vitro Methylation of Predetermined Regions in Recombinant DNA Constructs
- In Situ Detection of DNA Strand Breaks in Analysis of Apoptosis by Flow- and Laser-Scanning Cytometry
- Determination of Loss of Heterozygosity Using Polymerase Chain Reaction
- An Extended Suite of Genetic Tools for Use in Bacteria of the Halomonadaceae: An Overview
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