Genotyping and mapping assay of single-nucleotide polymorphisms in CYP3A5 using DNA-binding zinc(II) complexes
Clinical Biochemistry Volume 43 Issue 3
Page 302-306
published_at 2010-02
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| File | |
| Title ( eng ) |
Genotyping and mapping assay of single-nucleotide polymorphisms in CYP3A5 using DNA-binding zinc(II) complexes
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| Creator |
Kinoshita-Kikuta Emiko
Nakashima Hiromi
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| Source Title |
Clinical Biochemistry
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| Volume | 43 |
| Issue | 3 |
| Start Page | 302 |
| End Page | 306 |
| Abstract |
Objective: It is clinically important to detect the single-nucleotide polymorphism (SNP) of CYP3A5*3 (6986A>G) associated with enzymatic activity for drug metabolism. The aim of this study was to establish an accurate strategy for SNP screening. Design and methods: Polyacrylamide gel electrophoresis (PAGE) using zinc(II) complexes were applied for SNP detection. Genomic analyses of 19 healthy subjects were conducted by using both Zn2+-Phos-tag-PAGE and Zn2+-cyclen-PAGE methodologies. Results: Zn2+-Phos-tag PAGE permitted identification of the following allele genotypes: the A/A homozygote (CMA5*1/*1) in 3 individuals, the G/G homozygote (CMA5*3/*3) in 14 individuals, and the A/G heterozygote (CMA5*1/*3) in 2 individuals. Zn2+-cyclen PAGE demonstrated not only reproducibility of the genotyping but also existence of a novel heterozygous SNP (6929G>A) in the subject with CMA5*1/*1. Conclusion: We demonstrated reliable SNP genotyping and mapping in CYP3A5 using the combination method of Zn2+-Phos-tag PAGE and Zn2+-cyclen PAGE.
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| Keywords |
CYP3A5
SNP
Mutation
Zinc(II) complex
Phos-tag
Cyclen
PAGE
Electrophoresis
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| NDC |
Biology [ 460 ]
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| Language |
eng
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| Resource Type | journal article |
| Publisher |
Pergamon
Elsevier Science Ltd
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| Date of Issued | 2010-02 |
| Rights |
Copyright (c) 2009 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
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| Publish Type | Author’s Original |
| Access Rights | open access |
| Source Identifier |
[ISSN] 0009-9120
[DOI] 10.1016/j.clinbiochem.2009.09.007
[NCID] AA00607603
[DOI] http://dx.doi.org/10.1016/j.clinbiochem.2009.09.007
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