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?

***** 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 ***** Clostridium difficile toxins. Second, the authors also wanted to show that ***** destabilizes the TcdR-containing holoenzyme. This further suggests ***** TcdC has an ability ***** regulate toxin expression in its unique way. To understand the relevance of ***** two objectives incited, 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 other *****s such as TcdA, TcdB, and TcdR, genes that positively regulate the production ***** toxins A and B. It has *****en postulated that TcdC negatively regulates ***** gene expression of these toxin-producing ***** (Govind et al. 2006; Matamorous et al 2007). TcdC ***** highly expressed when C. difficile are found to grow exponenti*****ly, and expression is silenced ***** cells enter the stationary phase; TcdC expression is upped when ***** toxin-producing *****s are shut **********. There have ***** been studies that demonstrated mutations in the TcdC genes among epidemic strains ***** produce both Toxins A ***** ***** (Chernak and Johnson 2005; Matamorous et al. 2007).

Evidence to support ***** ***** aforementioned problems ***** 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 toxins produced by *****, TcdB, and TcdR genes. There has also ***** a growing number of ***** that supports the presence of Tcd***** ***** among epidemic strains of C. difficile. To prove that Tcd***** negatively ***** Tcd*****, TcdB and ***** expression may allow a more in-depth understanding of the pathogenesis of C. difficile. In turn, this may be a good foundation in developing medications ***** could combat ***** effects ***** this ***** pathogenic microbe. An understanding of the negative regulatory ***** ***** TcdC can help in addressing epidemic outbreaks.

Is the design of the experiment appropriate and efficient for the question? How else the experiment might have been designed and would an alternate design be better or worse?

***** quantitative, experimental study essentially had two major parts. First, the authors tested ***** ability of ***** to repress TcdA *****. This was demonstrated by using in vivo and ***** 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***** activity in ***** absence of TcdR demonstrated that there was a consequent lack ***** tcdA expression. This was in contrast ***** the ********** in b-glucuronidase ***** with TcdR. However, in the culture with an overexpression of Tcd*****, no glucuronidase activity was demonstrated despite the presence of *****R. *****s ***** to repress


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