UNDER THE SUPERVISION PRESENTED BY

OF
M.Sc.-4th SEM.,(BOTANY).
Reverse transcription is the process in which a
double stranded DNA molecules are made from a
single stranded RNA.

The name of this method is formed by its opposite

direction to transcription.
It also involves the presence of a reverse
transcriptase enzyme, a primer [18 - 22 bases nucleotide
segments], and a RNase inhibitor (RI). [leucine rich repeat
protein which binds with certain RNase with the highest affinity for
any protein-protein interaction].

RNase inhibitor is used in most laboratories

working with RNA to sequester RNase and, thereby,
protect the RNA in their samples from degradation
of by environmental RNase.
RNase is an enzyme that degrades RNA molecules
into smaller products. (i.e.- oligoribonucleotide and
ribonucleotide).

A reverse transcriptase, also known as RNA-

dependent DNA polymerase, is a DNA polymerase
enzyme that transcribes single-stranded RNA into
double-stranded DNA.
Normal transcription involves the synthesis of RNA
from DNA; hence, reverse transcription is the reverse
of this.

Reverse Transcriptase was discovered by Howard

Temin and Baltimore in 1970 independently.

Shared Nobel Prize in Physiology or Medicine in

1975 for their discovery.
The other names of Reverse Transcriptase is RNA-
directed DNA Polymerase, DNA Nucleotidyl
transferase (RNA directed), or Revertase.

Reverse transcriptase common in Retrovirus.

Reverse Transcriptase have fallowing activity:

1- RNA dependent DNA polymerase.
2- DNA dependent DNA polymerase.
3- RNase H.
 After the RNA retrovirus enters a host cell, its
genomic RNA will be transcribed into a double
stranded DNA and then integrated into the host
DNA. The RNA to DNA transcription is
called reverse transcription.
Figure - Mechanism of reverse transcription. The entire process is catalyzed
by reverse- transcriptase which has both DNA polymerase and RNase H
activities.
1. A retrovirus-specific cellular tRNA hybridizes with a
complementary region called the primer-binding site (PBS).

2. A DNA segment is extended from tRNA based on the sequence

of the retroviral genomic RNA.

3. The viral R and U5 sequences are removed by RNase H.

4. First jump: DNA hybridizes with the remaining R sequence at

the 3' end.

5. A DNA strand is extended from the 3' end.

6. Most viral RNA is removed by RNase H.

7. A second DNA strand is extended from the viral RNA.

8. Both tRNA and the remaining viral RNA are removed by

RNase H.

9. Second jump: The PBS region of the second strand hybridizes

with the PBS region of the first strand.

10. Extension on both DNA strands. LTR stands for "long

terminal repeat".
A. In 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 The molecular structure
to disrupt the process and thereby of zidovudine (AZT), a drug
used to inhibit HIV reverse
suppress its growth.
transcriptase.
 Collectively, these drugs are known as reverse-
transcriptase inhibitors and include the nucleoside and
nucleotide analogues zidovudine (trade name Retrovir)
, lamivudine (Epivir) and tenofovir (Viread) , as well as
non-nucleoside inhibitors, such as nevirapine (Viramune).

B. In Molecular biology and recombinant

DNA technology
1- cDNA formation -
 Reverse transcriptase is used to obtain DNA copies of RNA
molecules. i.e., cDNAs (complementary DNA or copy DNA)
Figure - Production of cDNA from an mRNA molecules used as
template by the enzyme reverse transcriptase.
 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.
Figure - RT - PCR
 Reverse transcriptase is used also to create cDNA
libraries from mRNA.

 Reverse transcriptase has also been employed

in insulin production.

 The commercial availability of reverse transcriptase

greatly improved knowledge in the area of molecular
biology, as, along with other enzymes, it allowed
scientists to cloning, sequencing, and characterise
RNA.
1. B.D. SINGH, BIOTECHNOLOGY.
2. WIKIPEDIA, THE FREE ENCYCLOPEDIA.