Translation bacterial

The tasks of transcriptional and translational signal recognition involve the prediction of promoters and sites that function in the initiation and termination of transcription and translation. Bacterial promoter sites, specifically the Escherichia coli RNA polymerase promoter site, are now very well characterized. The main problem is that the two conserved regions of the bacterial promoter, the -10 and -35 regions, are separated from each other by 15 to 21 bases, making the detection of the entire promoter as a single pattern difficult. Eukaryotic promoters are less well characterized than their bacterial equivalents. The major elements are the CCAAT box, GC box, TATA box and cap site. 

Good L., Nielsen P.E. Inhibition of translation and bacterial growth by peptide nucleic acid targeted to ribosomal RNA. Proc. Natl Acad. Sci. USA 1998 95 2073-2076. 

Independent bacterial motion is a true movement of translation and must be distinguished from the quivering or back-and-forth motion exhibited by very small particles suspended in a liquid. This latter type of motion is called Brownian movement and is caused by the bombardment of the bacteria by the molecules of the suspending fluid. 

This chapter presents methods and protocols suitable for the identification and characterization of inhibitors of the prokaryotic and/or eukaryotic translational apparatus as a whole or targeting specific, underexploited targets of the bacterial protein synthetic machinery such as translation initiation and amino-acylation. Some of the methods described have been used successfully for the high-throughput screening of libraries of natural or synthetic compounds and make use of model universal mRNAs that can be translated with similar efficiency by cellfree extracts of bacterial, yeast, and HeLa cells. Other methods presented here are suitable for secondary screening tests aimed at identifying a ... 

The activity of the bacterial translational apparatus can be studied in cellfree systems programmed, depending upon the particular experimental need and design, with any of the mRNAs shown in Fig. 12.1 and listed in Table 12.1. The amount of synthesized product can be assessed using either a radioactive test or, when translation is directed by 027IFCp(A), an immunological test (see later). 

This is a secondary test, the purpose of which is to ascertain that translational inhibitors active on the yeast and/or bacterial translational apparatus are harmless for the human protein synthetic machinery. All the considerations made for the yeast translation apply also to this system. 

Translation extracts Rabbit reticulocyte lysates (RRL) (Promega), wheat germ (WG) extract (Promega), bacterial S30 extract (Promega). Extracts from Krebs-2 cells were prepared as described (Svitkin and Sonenberg, 2004). 

A number of stimuli are known to act as inducers of TNF production (Table 9.6). Bacterial LPS represents the most important inducer, and TNF mediates the pathophysiological effects of this molecule. TNF biosynthesis is regulated by both transcriptional and post-transcriptional mechanisms. Macrophages appear to express TNF-a mRNA constitutively, which is translated only... 

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