Essay - Molecular Biology Clostridium Difficile How Important is the Experimental Question...

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Molecular Biology Clostridium difficile

How important is the experimental question being addressed to the field?

The study of Matamorous, England, and Dupuy (2007) aimed to present two things. First, ***** authors would like to provide evidence that TcdC negatively regulates the synthesis of Clostridium difficile toxins. Second, the authors also wanted to show that ***** destabilizes the TcdR-containing holoenzyme. This further suggests that TcdC has an ability ***** regulate toxin expression in its unique way. To understand the relevance of ***** two objectives **********, it must first be established what TcdC is. TcdC gene ***** found at the right end of the pathogenicity locus (PaLoc) of C. difficile. It is found along with o*****r *****s such as TcdA, TcdB, and TcdR, genes that positively regulate the production of toxins A ***** B. It has *****en postulated that TcdC negatively regulates the gene expression of these toxin-producing *****s (Govind et al. 2006; Matam*****ous et ***** 2007). ***** is highly expressed when C. difficile are found to grow exponentially, and expression is silenced when cells enter the stationary p*****e; TcdC expression ***** upped when other ***** genes are shut **********. There have ***** been studies that demonstrated mutations in the TcdC genes among epidemic strains that produce both Toxins A and B (Chernak and Johnson 2005; Matamorous et al. 2007).

Evidence to support ***** ***** aforementioned problems is relevant ***** the field of molecular ***** clinical biology. TcdC is a rel*****tively new discovery. It is a membrane-associated protein with a relatively unknown regulatory mechanism. The virulence of C. difficile is due to the synthes***** of toxins produced by *****, TcdB, and TcdR genes. There has also been a *****ing number of evidence th*****t supports the presence of TcdC mutations among epidemic strains of C. difficile. To prove that ***** negatively regulates Tcd*****, *****B and TcdR expression may allow a more in-depth underst*****ing of the pathogenesis of *****. difficile. In turn, this may be a good foundation in developing medications ***** could combat ***** effects of this ***** pathogenic microbe. An understanding of the negative regulatory effects ***** TcdC can help in addressing epidemic outbreaks.

Is the design of ***** experiment appropriate and efficient for the *****? How else the experiment might have been designed ***** would an alternate design be b*****ter or worse?

***** quantitative, *****al study essentially had two major parts. First, the authors tested ***** ability ***** TcdC to repress TcdA *****. This was demonstrated by using in vivo and in vitro models. TcdC *****ive activity ***** compared to TcdR's augmentative effect in the expression of TcdA. Using C. perfingens to take the place of C. *****, β-glucuronidase activity was used to measure promoter activity; the lack of β-glucuronidase activity in the absence of TcdR demonstrated that there was a consequent lack of tcdA expression. This was in contrast ***** the *****crease in b-glucuronidase activity with TcdR. However, in the culture with an overexpression of Tcd*****, no glucuronidase activity was ***** despite the ***** of TcdR. *****s ***** to repress


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