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 ***** synthesis of Clostridium ***** *****xins. Second, the authors also wanted to show that ***** destabilizes ***** 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 *****cited, it must first be established what TcdC is. TcdC gene ***** found at the right end ***** ***** pathogenicity locus (PaLoc) of C. difficile. It is ***** along with other *****s such as TcdA, TcdB, and TcdR, genes that positively regulate the production of *****s A and B. It has been postulated that TcdC negatively ***** the gene expression of these toxin-producing genes (Govind et al. 2006; Matamorous et al 2007). TcdC is 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 ***** ***** genes are shut off. There have also been studies that demonstrated mutations in the TcdC genes among epidemic strains that produce both *****xins ***** and B (Chernak and Johnson 2005; Matamorous et al. 2007).

Evidence to support ***** two aforementioned problems is relevant to the field of molecular ***** clinical biology. TcdC is a rel*****tively new discovery. It is a membr*****ne-associated protein with a relatively unknown regula*****ry mechanism. The virulence of C. ***** is due to the synthesis of ********** produced by TcdA, Tcd*****, and TcdR *****. There has also been a *****ing number of evidence ***** supports the presence of TcdC ***** among epidemic strains of C. difficile. To prove that ***** negatively regulates TcdA, TcdB and ***** expression may allow a more in-depth understanding of the pathogenesis of *****. difficile. In turn, this may be a good foundation in developing medications ***** could combat ***** effects ***** this ***** pathogenic microbe. An ***** 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 ***** been designed ***** ***** an alternate design be b*****ter or w*****se?

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


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