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Final Exam

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B- hemolysin antibody is a polyclonal antibody, has a wide range of affinities and specificities and thus it can detect any antigen present in its environment.d) In anti-b blot, protein A produces a stronger signal for arg-a42 and no signal in arg-a40. The signal for B-hemolysin is present only in arg-a40 and not arg-a42. These results mean that different regions of P3 codes for different chemicals. The author used RN7220 because it can increase the hla (a- hemolysin coding sequence) and hlb (B-hemolysin coding sequence) transcripts while suppressing spa transcripts. RN7220 has RNAIII clone a27 (pRN6735) as its genotype. From the fusion experiments, the researchers learned that B-lactamase expression is controlled by arg since fusion produced B-lactamase expression only in arg+ strains but not arg- strains. Besides, the results also proved that arg does not have any effect on the mRNA stability but it acts through regulation of the transcription process. a) Transcription fusion is a technique that aims at measuring the transcription activity of the promoter by tagging. While, translation fusion allows to you follow up, locate and note the expression of a tagged protein.b) The hollow ones represent RN6390B strain, and the filled ones represent ISP546. It is because we noted earlier on that the a27 clone promotes hla transcripts, and it decreases the spa transcript in arg+. From our graphs, the hollow ones have increased more than the filled ones in a-hemolysin and vice versa in protein A.c) The conclusion was reached at due to the study of subclones in which some regions of RNAIII have been deleted and in some, the reading frames were inactivated by mutagenesis of specific sites on RNAIII. It was proven that despite the deletions the 3subclones like a39 and a28 still activate hla and hlb transcription more strongly to get to maximum activity at a length of 422 nt. Besides the 5 subclones also caused hla and hlb transcription. RNAIII regulate hla translation and transcription independently. 3 subclones like a28, a39, and a41, cause expression of hlb but not hla. It shows that hla mRNA translation does not take place in these strains with 3 subclones. However a-hemolysin expresses itself in strains with 5 subclone, a29. It shows that the translation of mRNA cannot happen in absence 5 region of RNAHI. a) The mechanism of actions proposed was that, RNAIII is
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