To investigate the efficiency of surveyor nuclease in error correction during gene synthesis. | [Annals of the Pakistan Institute of Medical Sciences • 2015]

Author(s):

1. Zia-Ur-Rehman : School of Applied Biosciences, National University of Sciences and Technology (NUST), Islamabad, Pakistan

2. Atta-Ur-Rahman : School of Applied Biosciences, National University of Sciences and Technology (NUST), Islamabad, Pakistan

3. Andrew Curin: Manchester Institute of Biotechnology, University of Manchester, UK

4. Lmran Tipu: Manchester Institute of Biotechnology, University of Manchester, UK

5. Philip J. R. Day: Manchester Institute of Biotechnology, University of Manchester, UK

6. Najam-Us-Sahar : School of Applied Biosciences, National University of Sciences and Technology (NUST), Islamabad, Pakistan

7. Sadaf Zaidi: School of Applied Biosciences, National University of Sciences and Technology (NUST), Islamabad, Pakistan

Abstract:

To establish the efficacy of Surveyor Nuclease in gene synthesis process Introduction: Synthetic biology provides tools to design "for the purpose" building of genome/gene and organisms. Approaches have adopted for the construction of different viral genomes like polio, etc. Error prone amplification of gene is the main hurdle in gene synthesis. The error may include insertion, deletion, substitution, etc. These error mainly arise from incorrect chemical synthesis of oligo,s or polymerase induced error. Therefore a sophisticated/efficient error correction method can give the better output of desired function of resulting protein. Materials and Methods: In this study the efficiency of Surveyor enzyme (a mismatch repair enzyme) was analysed on 611 by long hepatitis C virus (HCV) core gene. The synthesised core gene was cloned into pCDNA3.1+, transformed in DH5a and after mini prep sent for the DNA sequencing. Results: The result implicated that Surveyor enzyme showed maximum efficacy at 60min incubation. It can produce 60 per cent more error free gene as compared to Surveyor enzyme untreated genes. Conclusion: This method can reduce the errors upto 60 %, when this method is utilised for gene synthesis

Page(s): 176-179

DOI: DOI not available

Published: Journal: Annals of the Pakistan Institute of Medical Sciences, Volume: 11, Issue: 4, Year: 2015

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