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?

***** 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 of Clostridium ***** *****xins. 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 underst*****d the relevance of the two objectives incited, it must first be established what TcdC is. TcdC gene ***** found at the right end ***** the 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 ***** *****s A and B. It has been postulated that TcdC negatively regulates the gene expression of these toxin-producing *****s (Govind et al. 2006; Matamorous et ***** 2007). TcdC is highly expressed when C. difficile are found to grow exponentially, and expression is silenced ***** cells enter the stationary phase; ***** expression ***** upped when ***** toxin-producing genes are shut off. There have ***** been studies that demonstrated mutations in the TcdC genes among epidemic strains ***** produce both *****xins ***** and ***** (Chernak and Johnson 2005; ***** et al. 2007).

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

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

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


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