A A A A A A 3' 5° MRNA Add a poly-dT primer that binds to the polyA tail of MRNA. 3'1 :5' тт A A A A A A 5° :3" Add reverse transcriptase + DNTPS to synthesize a complementary DNA strand. 3' :5' ттT A A A A A A 5° 3' Add RNaseH to cut up the RNA and generate RNA primers. 3' 5' Add DNA polymerase I and DNA ligase to synthesize the second DNA strand. 3' 5' ÄÄÄÄ Ä Ä ШШ 5" :3' Double-stranded CDNA FIGURE 21.3 Synthesis of cDNA. A poly-dT primer binds to the 3' end of eukaryotic mRNAs. Reverse transcriptase catalyzes the synthesis of a complementary DNA strand (CDNA). RNaseH digests the MRNA into short pieces that are used as primers by DNA polymerase I to synthesize the second DNA strand. The 5' to 3' exonuclease function of DNA polymerase I removes all of the RNA primers except the one at the 5' end (because there is no primer upstream from this site). This RNA primer can be removed by the subsequent addition of an RNase. After the double-stranded CDNA is made, it can then be inserted into a vector, as described in Figure 21.4b.
A A A A A A 3' 5° MRNA Add a poly-dT primer that binds to the polyA tail of MRNA. 3'1 :5' тт A A A A A A 5° :3" Add reverse transcriptase + DNTPS to synthesize a complementary DNA strand. 3' :5' ттT A A A A A A 5° 3' Add RNaseH to cut up the RNA and generate RNA primers. 3' 5' Add DNA polymerase I and DNA ligase to synthesize the second DNA strand. 3' 5' ÄÄÄÄ Ä Ä ШШ 5" :3' Double-stranded CDNA FIGURE 21.3 Synthesis of cDNA. A poly-dT primer binds to the 3' end of eukaryotic mRNAs. Reverse transcriptase catalyzes the synthesis of a complementary DNA strand (CDNA). RNaseH digests the MRNA into short pieces that are used as primers by DNA polymerase I to synthesize the second DNA strand. The 5' to 3' exonuclease function of DNA polymerase I removes all of the RNA primers except the one at the 5' end (because there is no primer upstream from this site). This RNA primer can be removed by the subsequent addition of an RNase. After the double-stranded CDNA is made, it can then be inserted into a vector, as described in Figure 21.4b.
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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Question
Explain the meaning of the name reverse transcriptase.
Transcribed Image Text:A A A A A A
3'
5°
MRNA
Add a poly-dT primer that binds
to the polyA tail of MRNA.
3'1
:5'
тт
A A A A A A
5°
:3"
Add reverse transcriptase
+ DNTPS to synthesize a
complementary DNA strand.
3'
:5'
ттT
A A A A A A
5°
3'
Add RNaseH to
cut up the RNA
and generate
RNA primers.
3'
5'
Add DNA polymerase I and
DNA ligase to synthesize
the second DNA strand.
Transcribed Image Text:3'
5'
ÄÄÄÄ Ä Ä
ШШ
5"
:3'
Double-stranded CDNA
FIGURE 21.3 Synthesis of cDNA. A poly-dT primer binds to
the 3' end of eukaryotic mRNAs. Reverse transcriptase catalyzes the
synthesis of a complementary DNA strand (CDNA). RNaseH digests the
MRNA into short pieces that are used as primers by DNA polymerase I
to synthesize the second DNA strand. The 5' to 3' exonuclease function
of DNA polymerase I removes all of the RNA primers except the one at
the 5' end (because there is no primer upstream from this site). This RNA
primer can be removed by the subsequent addition of an RNase. After
the double-stranded CDNA is made, it can then be inserted into a vector,
as described in Figure 21.4b.
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