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

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

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

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


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