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 ***** *****xins. 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 **********, it must first be established what ***** is. TcdC gene is 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 ***** *****s A and B. It has *****en postulated that TcdC negatively regulates the gene expression of 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 when other toxin-producing genes are shut off. There have ***** been studies that demonstrated mutations in the TcdC genes among epidemic strains that produce both *****xins A ***** B (Chernak and Johnson 2005; ***** et al. 2007).

Evidence to support the two 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 rel*****tively unknown regula*****ry mechanism. The virulence of C. difficile is due to the synthes***** of ********** produced by *****, TcdB, and TcdR genes. There has also ***** a *****ing number of ***** ***** supports the presence of ***** mutations among epidemic strains of *****. difficile. To prove that TcdC negatively ***** 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 **********tion in developing medications ***** could combat the effects of this highly 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 ***** might have been designed ***** ***** an alternate design be better or worse?

This quantitative, experimental study essentially had two major parts. First, the authors tested ***** ability of ***** to repress ***** expression. This was demonstrated ***** using in vivo and in vitro models. TcdC repressive activity was comp*****d 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 the absence ***** TcdR demonstrated that *****re ***** a consequent lack of tcdA expression. This was in contrast to the increase in b-glucuronidase ***** with TcdR. However, in the culture ***** an overexpression of Tcd*****, no glucuronidase activity was demonstrated despite the ***** of TcdR. *****s ability ***** repress


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