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Reverse Transcriptase (RNA-dependent DNA Polymerase)

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Presentation on theme: "Reverse Transcriptase (RNA-dependent DNA Polymerase)"— Presentation transcript:

1 Reverse Transcriptase (RNA-dependent DNA Polymerase)
Crystallographic structure of reverse transcriptase

2 RNA dependent DNA Polymerase
In the fields of molecular biology and biochemistry, a reverse transcriptase, also known as RNA dependent DNA polymerase, is a DNA polymerase enzyme that transcribes single stranded RNA into double-stranded DNA. It also helps in the formation of a double helix DNA once the RNA has been reverse transcribed into a single strand cDNA. Normal transcription involves the synthesis of RNA from DNA; hence, reverse transcription is the reverse of this.

3 Functions in virus The enzyme is encoded and used by reverse-transcribing viruses, which use the enzyme during the process of replication. Reverse-transcribing RNA viruses, such as retroviruses, use the enzyme to reverse-transcribe their RNA genomes into DNA, which is then integrated into the host genome and replicated along with it. HIV infects humans with the use of this enzyme. Without reverse transcriptase, the viral genome would not be able to incorporate into the host cell, resulting in the failure of the ability to replicate.

4 Process of Replication
Reverse transcriptase creates single stranded DNA from an RNA template. The process of reverse transcription is extremely error-prone and it is during this step that mutations may occur. Such mutations may cause drug resistance

5 Antiviral Drugs As HIV uses reverse transcriptase to copy
its genetic material and generate new viruses (part of a retrovirus proliferation circle), specific drugs have been designed to disrupt the process and thereby suppress its growth. Collectively, these drugs are known as reverse transcriptase inhibitors.

6 Applications of Reserve transcriptase in Molecular Biology
Reverse transcriptase is commonly used in research to apply the polymerase chain reaction technique to RNA in a technique called reverse transcription polymerase chain reaction (RT-PCR). The classical PCR technique can be applied only to DNA strands, but, with the help of reverse transcriptase, RNA can be transcribed into DNA, thus making PCR analysis of RNA molecules possible. Reverse transcriptase is used also to create cDNA libraries from mRNA. The commercial availability of reverse transcriptase allows scientists to clone, sequence, and characterize DNA. Reverse transcriptase has also been employed in insulin production. By Inserting eukaryotic mRNA for insulin production along with reverse transcriptase into bacteria, the mRNA can insert itself into the bacteria’s genome, and large amounts of insulin can be created, decreasing the need to harvest pig pancreas and other such traditional sources. Inserting eukaryotic DNA (instead of mRNA) into Bacteria would not work because it is fragmented, with introns, and would not transcribe successfully using the bacteria's ribosome.

7 QUIZ What is a reverse transcriptase? a) a DNA dependent polymerase
b) a RNA –dependent DNA Polymerase c) a RNA dependent Polymerase d) a virus reverse the transcription process

8 QUIZ 2) A reverse transcriptase is a (an) a) protein b) virus
c) nucleic acid d) fatty acid

9 QUIZ A reverse transcriptase has how many subunits? a) one b) two
c) three d) four

10 QUIZ The function of the reverse transcriptase is to
a) to transcribe complimentary strand DNA into RNA b) to cleave the double strand DNA into single strand DNA c) to transcribe single strand RNA into double stranded DNA d) to make copies of more single strand RNA

11 QUIZ The reverse transcriptase is used during the process of:
a) replication b) translation c) transcription d) cell division

12 Challenge Question If you are a scientist and want to develop a
drug to inhibit viral infection, what do you need to know to accomplish this mission?

13 Answers 1) b 2) a 3) b 4) c 5) c

14 Answer to Challenge Question
Answer may vary between students. The main idea is that students need to understand the pathway how the virus infect the host cell, what enzymes are required in the process and what drugs can be developed to prevent the enzymes formation, thus prevent the lifecycle of the virus to continue.


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