Use of megaprimer and overlapping extension pcr (OE-PCR) to mutagenize and enhance cyclodextrin glucosyltransferase (CGTASE) function
Protein engineering is a very useful tool for probing structure–function relationships in proteins. Specifically, site-directed mutagenized proteins can provide useful insights into structural, binding and catalytic mechanisms of a protein, particularly when coupled with crystallization. In this cha...
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| Format: | Book Section |
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Humana Press Inc.
2017
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| _version_ | 1796866344480669696 |
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| author | Goh, Kian Mau Liew, Kok Jun Chai, Kian Piaw Md. Illias, Rosli |
| author_facet | Goh, Kian Mau Liew, Kok Jun Chai, Kian Piaw Md. Illias, Rosli |
| author_sort | Goh, Kian Mau |
| collection | ePrints |
| description | Protein engineering is a very useful tool for probing structure–function relationships in proteins. Specifically, site-directed mutagenized proteins can provide useful insights into structural, binding and catalytic mechanisms of a protein, particularly when coupled with crystallization. In this chapter, we describe two protocols for performing site-directed mutagenesis of any protein-coding sequence, namely, megaprimer PCR and overlapping extension PCR (OE-PCR). We use as an example how these two SDM methods enhanced the function of a cyclodextrin glucosyltransferase (CGTase) from Bacillus lehensis strain G1.Bacillus, CGTase, Megaprimer PCR, Overlapping extension PCR, Protein engineering, Rational designProtein engineering is a very useful tool for probing structure–function relationships in proteins. Specifically, site-directed mutagenized proteins can provide useful insights into structural, binding and catalytic mechanisms of a protein, particularly when coupled with crystallization. In this chapter, we describe two protocols for performing site-directed mutagenesis of any protein-coding sequence, namely, megaprimer PCR and overlapping extension PCR (OE-PCR). We use as an example how these two SDM methods enhanced the function of a cyclodextrin glucosyltransferase (CGTase) from Bacillus lehensis strain G1. |
| first_indexed | 2024-03-05T21:10:44Z |
| format | Book Section |
| id | utm.eprints-97041 |
| institution | Universiti Teknologi Malaysia - ePrints |
| last_indexed | 2024-03-05T21:10:44Z |
| publishDate | 2017 |
| publisher | Humana Press Inc. |
| record_format | dspace |
| spelling | utm.eprints-970412022-09-15T04:16:35Z http://eprints.utm.my/97041/ Use of megaprimer and overlapping extension pcr (OE-PCR) to mutagenize and enhance cyclodextrin glucosyltransferase (CGTASE) function Goh, Kian Mau Liew, Kok Jun Chai, Kian Piaw Md. Illias, Rosli TP Chemical technology Protein engineering is a very useful tool for probing structure–function relationships in proteins. Specifically, site-directed mutagenized proteins can provide useful insights into structural, binding and catalytic mechanisms of a protein, particularly when coupled with crystallization. In this chapter, we describe two protocols for performing site-directed mutagenesis of any protein-coding sequence, namely, megaprimer PCR and overlapping extension PCR (OE-PCR). We use as an example how these two SDM methods enhanced the function of a cyclodextrin glucosyltransferase (CGTase) from Bacillus lehensis strain G1.Bacillus, CGTase, Megaprimer PCR, Overlapping extension PCR, Protein engineering, Rational designProtein engineering is a very useful tool for probing structure–function relationships in proteins. Specifically, site-directed mutagenized proteins can provide useful insights into structural, binding and catalytic mechanisms of a protein, particularly when coupled with crystallization. In this chapter, we describe two protocols for performing site-directed mutagenesis of any protein-coding sequence, namely, megaprimer PCR and overlapping extension PCR (OE-PCR). We use as an example how these two SDM methods enhanced the function of a cyclodextrin glucosyltransferase (CGTase) from Bacillus lehensis strain G1. Humana Press Inc. 2017 Book Section PeerReviewed Goh, Kian Mau and Liew, Kok Jun and Chai, Kian Piaw and Md. Illias, Rosli (2017) Use of megaprimer and overlapping extension pcr (OE-PCR) to mutagenize and enhance cyclodextrin glucosyltransferase (CGTASE) function. In: In Vitro Mutagenesis. Methods in Molecular Biology, 1498 (NA). Humana Press Inc., NA, pp. 385-396. ISBN 978-1-4939-6470-3 http://dx.doi.org/10.1007/978-1-4939-6472-7_27 DOI : 10.1007/978-1-4939-6472-7_27 |
| spellingShingle | TP Chemical technology Goh, Kian Mau Liew, Kok Jun Chai, Kian Piaw Md. Illias, Rosli Use of megaprimer and overlapping extension pcr (OE-PCR) to mutagenize and enhance cyclodextrin glucosyltransferase (CGTASE) function |
| title | Use of megaprimer and overlapping extension pcr (OE-PCR) to mutagenize and enhance cyclodextrin glucosyltransferase (CGTASE) function |
| title_full | Use of megaprimer and overlapping extension pcr (OE-PCR) to mutagenize and enhance cyclodextrin glucosyltransferase (CGTASE) function |
| title_fullStr | Use of megaprimer and overlapping extension pcr (OE-PCR) to mutagenize and enhance cyclodextrin glucosyltransferase (CGTASE) function |
| title_full_unstemmed | Use of megaprimer and overlapping extension pcr (OE-PCR) to mutagenize and enhance cyclodextrin glucosyltransferase (CGTASE) function |
| title_short | Use of megaprimer and overlapping extension pcr (OE-PCR) to mutagenize and enhance cyclodextrin glucosyltransferase (CGTASE) function |
| title_sort | use of megaprimer and overlapping extension pcr oe pcr to mutagenize and enhance cyclodextrin glucosyltransferase cgtase function |
| topic | TP Chemical technology |
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