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 the synthesis ***** Clostridium difficile toxins. Second, the authors also wanted to show that TcdC destabilizes ***** TcdR-containing holoenzyme. This further suggests ***** TcdC has an ability to 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 ***** 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 of *****s A and B. It has *****en postulated that TcdC negatively regulates ***** gene expression ***** these toxin-producing ***** (Govind et al. 2006; Matamorous et ***** 2007). TcdC is highly expressed when C. ***** are found to grow exponentially, and expression is silenced ***** cells enter the stationary p*****e; ***** expression is upped when other ***** *****s are shut off. There have ***** been studies that demonstrated mutations in the TcdC genes among epidemic strains ***** produce both Toxins A ***** ***** (Chernak and Johnson 2005; ***** et al. 2007).

Evidence to support ***** ***** aforementioned problems ***** relevant to the field of molecular and 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. difficile is due to the ***** of toxins produced by TcdA, TcdB, and TcdR *****. There has also ***** a growing number of ***** th*****t supports the presence of TcdC ***** among ***** strains of C. difficile. To prove that ***** 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 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 ***** experiment appropriate and efficient for the *****? How else the experiment might ***** been designed and would an alternate design be b*****ter or w*****se?

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


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