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 ***** field?

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

Evidence to support ***** ***** aforementioned problems ***** relevant to the field of molecular ***** clinical biology. TcdC is a rel*****tively new discovery. It is a membr*****ne-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 *****. There has also ***** a growing number of evidence ***** supports the presence of TcdC mutations among ***** strains of C. difficile. To prove that TcdC negatively regulates Tcd*****, *****B and TcdR expression may allow a more in-depth underst*****ing of the pathogenesis of C. difficile. In turn, this may be a good *****ation in developing medications ***** could combat ***** effects ***** this highly pathogenic microbe. An understanding of the negative regulatory ***** of TcdC can help in addressing epidemic outbreaks.

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

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


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