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 the field?

***** study of Matamorous, England, and Dupuy (2007) aimed to present two things. First, ***** authors would like to provide evidence that TcdC negatively regulates the synthesis of 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 ********** expression in its unique way. To understand the relevance of the two objectives incited, it must first be established what TcdC is. TcdC gene is found at the right end ***** ***** pathogenicity locus (PaLoc) of C. difficile. It is ***** along with other genes such as TcdA, TcdB, and TcdR, genes that positively regulate the production ***** toxins A ***** B. It has been postulated that TcdC negatively ***** the gene expression of these toxin-producing genes (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 phase; TcdC expression is upped when ***** ***** genes are shut off. There have ***** been studies that demonstrated mutations in the ***** genes among epidemic strains that produce both Toxins A ***** B (Chernak and Johnson 2005; ***** et al. 2007).

Evidence to support ***** two aforementioned problems ***** relevant to the field of molecular ***** clinical biology. TcdC is a relatively 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 ***** of toxins produced by TcdA, Tcd*****, and *****R genes. There has also been a growing number of ***** that supports the presence of Tcd***** mutations among epidemic strains of C. difficile. To prove that Tcd***** negatively regulates TcdA, TcdB and TcdR expression may allow a more in-depth underst*****ing of the pathogenes***** of C. difficile. In turn, this may be a good found*****tion in developing medications ***** could combat the effects of this highly pathogenic microbe. An understanding of the negative regulatory effects ***** TcdC can help in addressing epidemic outbreaks.

Is the design of ***** experiment appropriate and efficient for the question? How else the experiment might ***** been designed ***** ***** an alternate design be b*****ter or worse?

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

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