An Improved Technique for Genotyping the <i>ABCB1</i> Gene Variant of Exon 21
The Multidrug Resistance protein (<i>ABCB1</i>, <i>MDR1</i>) is involved in the transport of xenobiotics and antiretroviral drugs. Some variants of the <i>ABCB1</i> gene are of clinical importance; among them, exon 12 (c.1236C>T, rs1128503), 21 (c.2677G>T/A,...
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MDPI AG
2023-05-01
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| Online Access: | https://www.mdpi.com/2409-9279/6/3/53 |
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| author | Johanna Romina Zuccoli Priscila Ayelén Pagnotta Viviana Alicia Melito Jimena Verónica Lavandera Victoria Estela Parera Ana María Buzaleh |
| author_facet | Johanna Romina Zuccoli Priscila Ayelén Pagnotta Viviana Alicia Melito Jimena Verónica Lavandera Victoria Estela Parera Ana María Buzaleh |
| author_sort | Johanna Romina Zuccoli |
| collection | DOAJ |
| description | The Multidrug Resistance protein (<i>ABCB1</i>, <i>MDR1</i>) is involved in the transport of xenobiotics and antiretroviral drugs. Some variants of the <i>ABCB1</i> gene are of clinical importance; among them, exon 12 (c.1236C>T, rs1128503), 21 (c.2677G>T/A, rs2032582), and 26 (c.3435C>T, rs1045642) have a high incidence in Caucasians. Several protocols have been used for genotyping the exon 21 variants, such as allele-specific PCR-RFLP using adapted primer to generate a digestion site for several enzymes and automatic sequencing to detect the SNVs, TaqMan Allele Discrimination assay and High-Resolution Melter analysis (HRMA). The aim was to describe a new approach to genotype the three variants c.2677G>T/A for the exon 21 doing only one PCR with the corresponding primers and the digestion of the PCR product with two restriction enzymes: <i>BrsI</i> to identify A allele and <i>BseYI</i> to differentiate between G or T. An improvement of this methodology was also described. The proposal technique here described is demonstrated to be very efficient, easy, fast, reproducible, and cost-effective. |
| first_indexed | 2024-03-11T02:06:27Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2409-9279 |
| language | English |
| last_indexed | 2024-03-11T02:06:27Z |
| publishDate | 2023-05-01 |
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| series | Methods and Protocols |
| spelling | doaj.art-9c487c88d88b42849311454e0686d3032023-11-18T11:51:50ZengMDPI AGMethods and Protocols2409-92792023-05-01635310.3390/mps6030053An Improved Technique for Genotyping the <i>ABCB1</i> Gene Variant of Exon 21Johanna Romina Zuccoli0Priscila Ayelén Pagnotta1Viviana Alicia Melito2Jimena Verónica Lavandera3Victoria Estela Parera4Ana María Buzaleh5Centro de Investigaciones sobre Porfirinas y Porfirias (CIPYP), CONICET, Hospital de Clínicas José de San Martín, Universidad de Buenos Aires, Buenos Aires 1120, ArgentinaDepartamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires 2610, ArgentinaCentro de Investigaciones sobre Porfirinas y Porfirias (CIPYP), CONICET, Hospital de Clínicas José de San Martín, Universidad de Buenos Aires, Buenos Aires 1120, ArgentinaCátedra de Bromatología y Nutrición, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe 3000, ArgentinaCentro de Investigaciones sobre Porfirinas y Porfirias (CIPYP), CONICET, Hospital de Clínicas José de San Martín, Universidad de Buenos Aires, Buenos Aires 1120, ArgentinaCentro de Investigaciones sobre Porfirinas y Porfirias (CIPYP), CONICET, Hospital de Clínicas José de San Martín, Universidad de Buenos Aires, Buenos Aires 1120, ArgentinaThe Multidrug Resistance protein (<i>ABCB1</i>, <i>MDR1</i>) is involved in the transport of xenobiotics and antiretroviral drugs. Some variants of the <i>ABCB1</i> gene are of clinical importance; among them, exon 12 (c.1236C>T, rs1128503), 21 (c.2677G>T/A, rs2032582), and 26 (c.3435C>T, rs1045642) have a high incidence in Caucasians. Several protocols have been used for genotyping the exon 21 variants, such as allele-specific PCR-RFLP using adapted primer to generate a digestion site for several enzymes and automatic sequencing to detect the SNVs, TaqMan Allele Discrimination assay and High-Resolution Melter analysis (HRMA). The aim was to describe a new approach to genotype the three variants c.2677G>T/A for the exon 21 doing only one PCR with the corresponding primers and the digestion of the PCR product with two restriction enzymes: <i>BrsI</i> to identify A allele and <i>BseYI</i> to differentiate between G or T. An improvement of this methodology was also described. The proposal technique here described is demonstrated to be very efficient, easy, fast, reproducible, and cost-effective.https://www.mdpi.com/2409-9279/6/3/53<i>ABCB1</i> geneExon 21gene variantsgenotyping methodsingle nucleotide variantsrestriction fragment length polymorphisms |
| spellingShingle | Johanna Romina Zuccoli Priscila Ayelén Pagnotta Viviana Alicia Melito Jimena Verónica Lavandera Victoria Estela Parera Ana María Buzaleh An Improved Technique for Genotyping the <i>ABCB1</i> Gene Variant of Exon 21 Methods and Protocols <i>ABCB1</i> gene Exon 21 gene variants genotyping method single nucleotide variants restriction fragment length polymorphisms |
| title | An Improved Technique for Genotyping the <i>ABCB1</i> Gene Variant of Exon 21 |
| title_full | An Improved Technique for Genotyping the <i>ABCB1</i> Gene Variant of Exon 21 |
| title_fullStr | An Improved Technique for Genotyping the <i>ABCB1</i> Gene Variant of Exon 21 |
| title_full_unstemmed | An Improved Technique for Genotyping the <i>ABCB1</i> Gene Variant of Exon 21 |
| title_short | An Improved Technique for Genotyping the <i>ABCB1</i> Gene Variant of Exon 21 |
| title_sort | improved technique for genotyping the i abcb1 i gene variant of exon 21 |
| topic | <i>ABCB1</i> gene Exon 21 gene variants genotyping method single nucleotide variants restriction fragment length polymorphisms |
| url | https://www.mdpi.com/2409-9279/6/3/53 |
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