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

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

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

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


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