Nuclei have been isolated from a stem cell line at 0, 1, 2, and 4 days after the
ID: 215049 • Letter: N
Question
Nuclei have been isolated from a stem cell line at 0, 1, 2, and 4 days after the cell line has been induced to differentiate into muscle type cells. The nuclei are subjected to digestion with DNase, the DNA is then extracted from the nuclei and cleaved with a restriction enzyme that cleaves the DNA external to genes X and Y. The digestion products are run on a gel and analyzed by Southern blots using a labeled probe that binds to either X or Y DNA (or RNA). The results of this experiment are shown on the Southern blot below at the left. The experiment is repeated, but without DNase, and the results are shown on the Southern blot below in the middle. Finally, RNA, extracted from the cells at days 0, 1, 2 and 4 after induction, is run on gels which are then blotted and probed for the presence of X or Y RNA, as shown in the Northern blot below, right.
Is either gene X or gene Y likely to encode a muscle cell specific protein? Why?
Days of induction01 2 4 0 1 24 Probe X Probe Y | Southern blot Southern blot Northern blotExplanation / Answer
The northern blot analysis of cells indicates that cells which are not treated with DNAse continues the level of transcription and produce RNA. Induction of cells leads to increase in muscle RNA and decrease in other RNA which are not involved in muscle formation.
Southern blot first indicates that induction leads to decrease of Gene X and increase of gene Y.
but Southern blot to indicates that continuous expression of both DNA because here we do not used DNAse. Because DNAse denatures The DNA
Image 3 indicates that when we do not give any kind of DNAse enzyme treatment than DNA is completely able to produce high quantity of RNA. but due to induction the required RNA which is specific for muscle formation is increased which is by gene X and RNA which is not associated with specification of muscle cells and decreased. So that we can say that the X is associated with muscle forming proteins.
When we give external induction which are specific for muscle formation than all other genes become an activated or less transcribed and the gene required for muscle formation is highly activated due to induction and produce more RNA which are involved in high protein formation. Here, after induction for muscle formation mRNA of X gene was enhanced and mRNA for Y gene is reduced.
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