Amplification and cloning of taq gene from thermus aquaticus YT-1 strain /

Introduction: DNA polymerase is the crucial enzyme in polymerase chain reaction (PCR). The polymerase derived from Thermus aquaticus was found to be superior to the historical DNA polymerase of Escherichia coli, as it can withstand the extreme denaturation temperature during the PCR procedure. This...

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Bibliographic Details
Main Author: Elsalami, Rabia Mrehil Ali
Format: Thesis
Language:English
Published: Kuantan : Kulliyyah of Medicine, International Islamic University Malaysia, 2011
Subjects:
Online Access:Click here to view 1st 24 pages of the thesis. Members can view fulltext at the specified PCs in the library.
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040 |a UIAM  |b eng 
041 |a eng 
043 |a a-my--- 
050 |a RB43.8.P64 
100 1 |a Elsalami, Rabia Mrehil Ali 
245 1 |a Amplification and cloning of taq gene from thermus aquaticus YT-1 strain /  |c by Rabia Mrehil Ali ElSalami 
260 |a Kuantan :  |b Kulliyyah of Medicine, International Islamic University Malaysia,  |c 2011 
300 |a xii, 53 leaves :  |b ill. ;  |c 30cm. 
500 |a Abstract in English and arabic. 
500 |a "A dissertation submitted in fulfilment of the requirement for the degree of Master of Medical Sciences." --On t.p. 
502 |a Thesis (MMDSC)--International Islamic University Malaysia, 2011. 
504 |a Includes bibliographical references (leaves 49-51). 
520 |a Introduction: DNA polymerase is the crucial enzyme in polymerase chain reaction (PCR). The polymerase derived from Thermus aquaticus was found to be superior to the historical DNA polymerase of Escherichia coli, as it can withstand the extreme denaturation temperature during the PCR procedure. This study aims to amplify and clone the Taq gene from Thermus aquaticus YT-1 strain. Methods: Thermus aquaticus YT -1 strain was cultured on Castenholz medium. The bacterial colonies were then subjected to DNA extraction and purification processes to yield DNA for the PCR. The targeted fragment of Taq gene was amplified by using Hotstart PCR technique. The isolated amplified Taq gene was then digested with restriction enzymes and ligated into pUC18 cloning vector that had also been similarly cleaved. Following that the recombinant plasmid was transformed into ampicillin resistant Escherichia coli strain DH5a. PCR screening was done to confirm the insertion of Taq DNA gene. Results: the successful isolation of the Taq polymerase gene from Thermus aquaticus YT -1 strain was confirmed by DNA sequencing of the product. The study however failed to show a significant growth of transfom1ed E. coli colonies that carried the target gene. Discussion: even though the Taq polymerase gene was successfully isolated, the failure to successfully clone it into the cloning vector is speculated to be related to the relatively large size of the Taq gene that requires further optimization procedures to be perfonned on the employed methodology. Conclusion: the isolated Taq polymerase gene could be used as a foundation material for the development of further optimized and/or improvised gene cloning and expression techniques in a near future subsequent study. 
596 |a 1 
650 0 |a Polymerase chain reaction  |x Research 
650 0 |a DNA polymerases  |x Research 
655 7 |a Theses, IIUM local 
690 |a Dissertations, Academic  |x Department of Basic Medical Sciences  |z IIUM 
710 2 |a International Islamic University Malaysia.  |b Department of Basic Medical Sciences 
856 4 |u http://studentrepo.iium.edu.my/handle/123456789/5718  |z Click here to view 1st 24 pages of the thesis. Members can view fulltext at the specified PCs in the library. 
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