2. You have cloned the 4 bacterial species. Now you want to determine which one

Question

2. You have cloned the 4 bacterial species. Now you want to determine which one can degrade BA You therefore, mix pure BA with these bacteria and wait for a day. Then you measure the decrease in BA levels in each bacterial culture. You find out that species #3 (Sp3) reduces BA levels, and therefore, you decide that the gene which can degrade BA must be present in the genome of Sp.3. To find out which gene degrades BA, you make a microarray platform, with oligonucleotides corresponding to each gene of Sp.3. Then you perform the following experiment where you add the shown amounts of BA to Sp.3, and wait for the shown times, and you measure how much of BA is degraded. Results are shown in the below table. Experiment Nr BA added Time waited BA remaining at the end of experiment 10 12h 10 10 24h 0.0001 10 6h You then obtain mRNA from Sp.3 treated with BA for the times shown above and perform a microarray experiment. Some of the genes and the expression changes of these genes are shown below Time1h 6h 12h 24h Gene Nr 20 50 250 35 75 1000 37 500 100 250 1000 1500 35 Which of these 5 genes would you pick to study further (clone and purify) as a candidate BAse? Please explain your logic briefly

Explanation / Answer

2. Gene 4 would be picked up. The reason is simple, among the four experiment corresponding to 1, 6, 12 and 24 hrs, only in the fourth experiment i.e. at 24 hours does the BA levels decrease from 10 microgram to 0.0001 microgram. So the gene that is responsible for coding protein/enzyme to degrade BA must be expressed more than usual amounts in the fourth experiment. According to table 2, which shows the mRNA levels of all 5 genes in the four experiments, gene 4's expression increases exponentially with increase time in the four experiments and has the highest expression in the last experiment where the BA levels are reduced from 10 microgram to 0.0001 microgram. Thus, Gene 4 will be picked up for further studies.

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