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 difficile toxins. 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 understand the relevance of ***** two objectives *****cited, it must first be established what ***** is. TcdC gene ***** found at the right end of the pathogenicity locus (PaLoc) of C. difficile. It is found along with o*****r genes such as TcdA, TcdB, and TcdR, genes that positively regulate the production ***** ***** A and B. It has been postulated that TcdC negatively ***** the gene expression of these *****-producing genes (Govind et al. 2006; Matam*****ous et ***** 2007). ***** ***** highly expressed when C. ***** are found to grow exponentially, and expression is silenced when cells enter the stationary phase; TcdC expression is upped when other toxin-producing genes are shut off. There have also been studies that demonstrated mutations in the TcdC genes among epidemic strains ***** produce both *****xins A and B (Chernak and Johnson 2005; Matamorous et al. 2007).

Evidence to support ***** ***** 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, Tcd*****, and *****R genes. There has also ***** a *****ing number of evidence th*****t supports the presence of ***** mutations among epidemic strains of C. difficile. To prove that TcdC negatively regulates TcdA, TcdB and TcdR expression may allow a more in-depth underst*****ing of the pathogenesis of C. difficile. In turn, this may be a good found*****tion in developing medications ***** could combat the 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 have been designed and ***** an alternate design be b*****ter or w*****se?

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

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