Mutation detection in rice waxy mutants by PCR-RF-SSCP.
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| Title: | Mutation detection in rice waxy mutants by PCR-RF-SSCP. |
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| Authors: | Sato, Y.1, Nishio, T.1 nishio@bios.tohoku.ac.jp |
| Source: | Theoretical & Applied Genetics. Aug2003, Vol. 107 Issue 3, p560-567. 8p. |
| Subjects: | Plant genetic engineering, Genetic polymorphisms, Endonucleases, Messenger RNA, Rice, Plant genetics |
| Abstract: | PCR-RF-SSCP (PRS), which combines cleaved amplified polymorphic sequence (CAPS) and single-strand conformation polymorphism (SSCP), is expected to be a useful technique for DNA polymorphism analysis. We evaluated the ability of PRS to detect single nucleotide polymorphism (SNP) using the Waxy gene, Wx, of rice, and subsequently were able to identify point mutations in wx mutant lines. The approximately 6-kb Wx gene was divided into five regions for PCR amplification. Two regions, in which most of the point mutations of the wx mutants have been identified, were amplified by PCR and cloned into a vector, and those clones containing SNPs produced as a result of the inherent inaccuracy of PCR were used for the evaluation of PRS. The efficiency of PRS in the detection of SNPs of these clones was over 70%. PRS analysis of the wx genes in 18 waxy mutants was carried out in the five regions using two different restriction endonucleases and two gel conditions, with and without glycerol. Of the 18 lines tested, 17 showed band patterns different from that of the wild type. Most of the mutations identified in this study were nucleotide changes in exons, which result in amino acid changes. One mutation generated an in-frame stop codon, and another was a frame shift mutation by one-base deletion. Two mutations found at a splice site were considered to inhibit normal splicing of mRNA. These results show that PRS is a useful technique for detecting point mutations in large plant genes. [ABSTRACT FROM AUTHOR] |
| Copyright of Theoretical & Applied Genetics is the property of Springer Nature and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
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| Items | – Name: Title Label: Title Group: Ti Data: Mutation detection in rice waxy mutants by PCR-RF-SSCP. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Sato%2C+Y%2E%22">Sato, Y.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Nishio%2C+T%2E%22">Nishio, T.</searchLink><relatesTo>1</relatesTo><i> nishio@bios.tohoku.ac.jp</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Theoretical+%26+Applied+Genetics%22">Theoretical & Applied Genetics</searchLink>. Aug2003, Vol. 107 Issue 3, p560-567. 8p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Plant+genetic+engineering%22">Plant genetic engineering</searchLink><br /><searchLink fieldCode="DE" term="%22Genetic+polymorphisms%22">Genetic polymorphisms</searchLink><br /><searchLink fieldCode="DE" term="%22Endonucleases%22">Endonucleases</searchLink><br /><searchLink fieldCode="DE" term="%22Messenger+RNA%22">Messenger RNA</searchLink><br /><searchLink fieldCode="DE" term="%22Rice%22">Rice</searchLink><br /><searchLink fieldCode="DE" term="%22Plant+genetics%22">Plant genetics</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: PCR-RF-SSCP (PRS), which combines cleaved amplified polymorphic sequence (CAPS) and single-strand conformation polymorphism (SSCP), is expected to be a useful technique for DNA polymorphism analysis. We evaluated the ability of PRS to detect single nucleotide polymorphism (SNP) using the Waxy gene, Wx, of rice, and subsequently were able to identify point mutations in wx mutant lines. The approximately 6-kb Wx gene was divided into five regions for PCR amplification. Two regions, in which most of the point mutations of the wx mutants have been identified, were amplified by PCR and cloned into a vector, and those clones containing SNPs produced as a result of the inherent inaccuracy of PCR were used for the evaluation of PRS. The efficiency of PRS in the detection of SNPs of these clones was over 70%. PRS analysis of the wx genes in 18 waxy mutants was carried out in the five regions using two different restriction endonucleases and two gel conditions, with and without glycerol. Of the 18 lines tested, 17 showed band patterns different from that of the wild type. Most of the mutations identified in this study were nucleotide changes in exons, which result in amino acid changes. One mutation generated an in-frame stop codon, and another was a frame shift mutation by one-base deletion. Two mutations found at a splice site were considered to inhibit normal splicing of mRNA. These results show that PRS is a useful technique for detecting point mutations in large plant genes. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Theoretical & Applied Genetics is the property of Springer Nature and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1007/s00122-003-1282-4 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 8 StartPage: 560 Subjects: – SubjectFull: Plant genetic engineering Type: general – SubjectFull: Genetic polymorphisms Type: general – SubjectFull: Endonucleases Type: general – SubjectFull: Messenger RNA Type: general – SubjectFull: Rice Type: general – SubjectFull: Plant genetics Type: general Titles: – TitleFull: Mutation detection in rice waxy mutants by PCR-RF-SSCP. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Sato, Y. – PersonEntity: Name: NameFull: Nishio, T. IsPartOfRelationships: – BibEntity: Dates: – D: 01 M: 08 Text: Aug2003 Type: published Y: 2003 Identifiers: – Type: issn-print Value: 00405752 Numbering: – Type: volume Value: 107 – Type: issue Value: 3 Titles: – TitleFull: Theoretical & Applied Genetics Type: main |
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