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: | |
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| Format: | Article |
| Language: | English |
| Published: |
Pusat Pengajian Sains Perubatan, Universiti Sains Malaysia
2005
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| Subjects: | |
| 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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| Summary: | 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. |
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