g) You become highly ambitious at work and unfortunately you contaminate the E.
ID: 196940 • Letter: G
Question
g) You become highly ambitious at work and unfortunately you contaminate the E. palapyensis with E. gaboronensis. Because you have worked with the strains for long enough, you know that the two strains differ in a few bases on the promoter sequence as below: E. gaboronensis: CAGATACCATGGCTAAAAAGAAAATTCTGGCCGATTTTAGTCGT TGAAGAGCTTGCATGCATGCATGCATGCATGAATTC E. palapyensis: CAGATACCATGGCTAAAAAGAAAATTCTGGCCGATTTTAGTCGTTGA AAGCTTTGCATGCATGCATGCATGCATGAATTC i) After progressively showing how important you are to the company, your advisor suggests that you compare the levels of expression of the protein in both strains. ii. How would you separate the Lys mRNA from the total RNA?
Explanation / Answer
1. E. gaboronensis
CAG ATA CCA TGG CTA AAA AGA AAA TTC TGG CCG ATT TTA GTC GTT GAA GAG CTT GCA TGC ATG CAT GCA TGC ATG AAT TC
E. palapyensis:
CAG ATA CCA TGG CTA AAA AGA AAA TTC TGG CCG ATT TTA GTC GTT GAA AGC TTT GCA TGC ATG CAT GCA TGC ATG AAT TC
E. palapyensis has TATA box in the -10 promoter region. So, it express high level of protein compare to the E. gaboronensis which has no TATA box in -10 region.
2. Use poly dT beads to capture Lysine mRNAs since eukaryotic mRNA has poly-A tails and precipitate the other RNA using ethanol.
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