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, 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 *****xin expression in its unique way. To understand the relevance of ***** two objectives incited, it must first be established what ***** is. TcdC gene ***** found at the right end ***** ***** pathogenicity locus (PaLoc) of C. difficile. It is found along with other *****s such as TcdA, TcdB, and TcdR, genes that positively regulate the production of toxins A ***** B. It has been postulated that TcdC negatively ***** ***** gene expression ***** these toxin-producing *****s (Govind et al. 2006; Matamorous et al 2007). ***** ***** highly expressed when C. difficile are found to grow exponenti*****ly, and expression is silenced when cells enter the stationary p*****e; TcdC expression ***** upped ***** ***** ***** genes are shut off. There have ***** been studies that demonstrated mutations in the ***** genes among epidemic strains ***** produce both Toxins A and B (Chernak and Johnson 2005; Matamorous et al. 2007).

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

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

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


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