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Expression of recombinant thermus aquaticus dna polymerase gene

Taq DNA polymerase derived from the extreme thermophillic microorganism, Thermus aquaticus is very useful in Polymerase Chain Reaction (PCR) in which high temperature stable Deoxyribonucleotide acid (DNA polymerase is needed in amplifying a specific DNA fragment. It is also useful in DNA sequenc...

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Main Author: Thanabalan, Asha a/p
Format: Article
Language:English
Published: Pusat Pengajian Sains Perubatan, Universiti Sains Malaysia 2005
Subjects:
R Medicine
Online Access:http://eprints.usm.my/46973/1/PTA...Asha%20A%2CP%20Thanabalan...2005...mka..-24%20pages.pdf
http://eprints.usm.my/46973/
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spelling my.usm.eprints.46973 http://eprints.usm.my/46973/ Expression of recombinant thermus aquaticus dna polymerase gene Thanabalan, Asha a/p R Medicine Taq DNA polymerase derived from the extreme thermophillic microorganism, Thermus aquaticus is very useful in Polymerase Chain Reaction (PCR) in which high temperature stable Deoxyribonucleotide acid (DNA polymerase is needed in amplifying a specific DNA fragment. It is also useful in DNA sequencing. For this purpose, many techniques of cloning and expression of this enzyme were practiced to obtain a high performance enzyme. The purification methods were designed to retrieve a high enzyme yield. In this method, we have tried to express the recombinant Taq Pol I enzyme in different expression systems to select the best host for the protein expression. The host BL-21 was selected because of the additional property within it, a plysS plasmid for tighter protein expression but simpler purification method as adapted from Grimm et. al (1995). Protein expression was induced over a 12 hour period using IPTG before it was viewed in a large SDS- PAGE. There were bands along a marker indicating a complete protein expression as the weight of Taq DNA polymerase is 94 kD. Pusat Pengajian Sains Perubatan, Universiti Sains Malaysia 2005-03 Article NonPeerReviewed application/pdf en http://eprints.usm.my/46973/1/PTA...Asha%20A%2CP%20Thanabalan...2005...mka..-24%20pages.pdf Thanabalan, Asha a/p (2005) Expression of recombinant thermus aquaticus dna polymerase gene. Expression of recombinant thermus aquaticus dna polymerase gene. (Submitted)
institution Universiti Sains Malaysia
building Hamzah Sendut Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Sains Malaysia
content_source USM Institutional Repository
url_provider http://eprints.usm.my/
language English
topic R Medicine
spellingShingle R Medicine
Thanabalan, Asha a/p
Expression of recombinant thermus aquaticus dna polymerase gene
description Taq DNA polymerase derived from the extreme thermophillic microorganism, Thermus aquaticus is very useful in Polymerase Chain Reaction (PCR) in which high temperature stable Deoxyribonucleotide acid (DNA polymerase is needed in amplifying a specific DNA fragment. It is also useful in DNA sequencing. For this purpose, many techniques of cloning and expression of this enzyme were practiced to obtain a high performance enzyme. The purification methods were designed to retrieve a high enzyme yield. In this method, we have tried to express the recombinant Taq Pol I enzyme in different expression systems to select the best host for the protein expression. The host BL-21 was selected because of the additional property within it, a plysS plasmid for tighter protein expression but simpler purification method as adapted from Grimm et. al (1995). Protein expression was induced over a 12 hour period using IPTG before it was viewed in a large SDS- PAGE. There were bands along a marker indicating a complete protein expression as the weight of Taq DNA polymerase is 94 kD.
format Article
author Thanabalan, Asha a/p
author_facet Thanabalan, Asha a/p
author_sort Thanabalan, Asha a/p
title Expression of recombinant thermus aquaticus dna polymerase gene
title_short Expression of recombinant thermus aquaticus dna polymerase gene
title_full Expression of recombinant thermus aquaticus dna polymerase gene
title_fullStr Expression of recombinant thermus aquaticus dna polymerase gene
title_full_unstemmed Expression of recombinant thermus aquaticus dna polymerase gene
title_sort expression of recombinant thermus aquaticus dna polymerase gene
publisher Pusat Pengajian Sains Perubatan, Universiti Sains Malaysia
publishDate 2005
url http://eprints.usm.my/46973/1/PTA...Asha%20A%2CP%20Thanabalan...2005...mka..-24%20pages.pdf
http://eprints.usm.my/46973/
_version_ 1677782010038321152
score 13.211869

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