By Virginia L. Clark, Patrik M. Bavoil
The seriously acclaimed laboratory normal for greater than 40 years, Methods in Enzymology is without doubt one of the such a lot hugely revered guides within the box of biochemistry. due to the fact that 1955, each one quantity has been eagerly awaited, often consulted, and praised by way of researchers and reviewers alike. Now with greater than three hundred volumes (all of them nonetheless in print), the sequence includes a lot fabric nonetheless proper today—truly a vital e-book for researchers in all fields of existence sciences.
* provides possible choices to mammalian version systems
* Discusses virulence and crucial gene identification
* Defines international gene expression
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Additional resources for Bacterial Pathogenesis Part C: Identification, Regulation, and Function of Virulence Factors
C. elegans is an attractive model to study host defense response because the unprecedented combined power of genetic and functional genomic approaches can be applied to comprehensively dissect the innate immune system of a eukaryote host. In addition to its ease of genetic analysis, extensive molecular genetic toolbox, and fully sequenced genome, developments in rapid mapping, genome-wide expression using high-density arrays, and inhibition of gene function by RNA interference (RNAi) have made the genome-wide identification of host defense factors very accessible.
20 D. A. D'Argenio, L. A. Gallagher, C. A. Berg, and C. Manoil, J. Bacteriol. 183, 1466 (2001). 21 S. M. Wong andJ. J. Mekalanos, Proc. Natl. Acad. Sci. A. 97, 10191 (2000). ] Identification of Bacterial Mutants Defective in "Slow Killing. 'q5 Inoculate individual PA14::TnphoA clones into 200/zl King's B medium in microtiter plates containing rifampicin and neomycin at 100 and 200/zg/ml, respectively. 5-cm petri plates and incubate at 37 ° for at least 24 hr. With shorter incubation period, we often observe significant reduction in bacterial virulence.
Elegans Mutagenesis. EMS is the favored mutagen used in generating mutations in C. elegans because of its low toxicity and its ability to induce mutations at high frequencies. It is important to note that approximately 92% of EMSinduced base pair changes are G/C to A/T transitions, which favor the production of A/T-rich stop codon mutations but very rarely mutate amino acids Asp (AAY), lie (AUY, AUA), Lys (AAR), Phe (UUY), and Tyr (UAY) to other residues. 27 For genetic studies in which a variety of amino acid changes are desired, such as suppressor screens, N-ethyl-N-nitrosourea (ENU) is a superior mutagen because it does not produce the EMS-induced bias.
Bacterial Pathogenesis Part C: Identification, Regulation, and Function of Virulence Factors by Virginia L. Clark, Patrik M. Bavoil