What is the first step in quantifying the relative amounts of mRNA in different tissues? Would this method be useful in determining which immune system genes might be over-expressed in severe Covid cases? Why or why not?

Biochemistry
6th Edition
ISBN:9781305577206
Author:Reginald H. Garrett, Charles M. Grisham
Publisher:Reginald H. Garrett, Charles M. Grisham
Chapter29: Transcription And The Regulation Of Gene Expression
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  • What is the first step in quantifying the relative amounts of mRNA in different tissues? Would this method be useful in determining which immune system genes might be over-expressed in severe Covid cases? Why or why not?

  • Could quantitative PCR, which uses a DNA-binding dye, to show how many copies of the target DNA sequence could be used to quantify the amount of mRNA in a cell? Would you expect that a metabolically active tissue such as the liver would show more cDNA copies in such a method, compared to less metabolically active tissues such as skin cells?
  • One reason that the types and amounts of mRNAs are quantified in different tissue types is to compare which genes are activated and which are inactive.  It used to be thought that any gene that was transcribed was automatically translated. The discovery of RNA-degrading systems shows that the real situation in cells is more complemented. Do you believe that a larger amount of mRNA of a given type, say for alpha hemoglobin in immature red blood cells is a reliable indicator that more alpha hemoglobin protein will be made in those cells?

MRNA
S' m?G
AAAAAAAA 3'
oligo-dT primer is annealed
to MRNA template
5' m?G
AAAAAAAA 3'
TTTTTTTT 5'
Reverse transcriptase uses
DNTPS to synthesize a
complementary DNA strand
MRNA-DNA hybrid
S' m'G
AAAAAAAA 3'
TTTTTTTT 5'
MRNA is degraded with
alkali or RNase H
TTTTTTTT 5'
To prime synthesis of a
second strand, an
oligonucleotide of known
sequence is ligated to the
3' end of the CDNA
5'
3'
3'
тттттттт 5
DNA polymerase l and
DNTPS extend the primer to
yield double-stranded DNA
CDNA
5'
AAAAAAAA 3'
TTTTTTTT 5'
Figure 10-9
Introduction to Genetic Analysis, Twelfth Edition
O 2020 W. H. Freeman and Company
Transcribed Image Text:MRNA S' m?G AAAAAAAA 3' oligo-dT primer is annealed to MRNA template 5' m?G AAAAAAAA 3' TTTTTTTT 5' Reverse transcriptase uses DNTPS to synthesize a complementary DNA strand MRNA-DNA hybrid S' m'G AAAAAAAA 3' TTTTTTTT 5' MRNA is degraded with alkali or RNase H TTTTTTTT 5' To prime synthesis of a second strand, an oligonucleotide of known sequence is ligated to the 3' end of the CDNA 5' 3' 3' тттттттт 5 DNA polymerase l and DNTPS extend the primer to yield double-stranded DNA CDNA 5' AAAAAAAA 3' TTTTTTTT 5' Figure 10-9 Introduction to Genetic Analysis, Twelfth Edition O 2020 W. H. Freeman and Company
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