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Cloning flanking sequence by single-primer PCR in transgenic plants.
Ma, J; Wang, Y P; Ren, S; Zhang, Z; Lu, S; Wang, P W.
Afiliação
  • Ma J; Biotechnology Center of Jilin Agricultural University, Changchun, Jilin Province, China.
  • Wang YP; Biotechnology Center of Jilin Agricultural University, Changchun, Jilin Province, China.
  • Ren S; Jilin Provincial Center for Instruction of Undergraduates, Changchun, Jilin Province, China.
  • Zhang Z; Biotechnology Center of Jilin Agricultural University, Changchun, Jilin Province, China.
  • Lu S; Biotechnology Center of Jilin Agricultural University, Changchun, Jilin Province, China.
  • Wang PW; Biotechnology Center of Jilin Agricultural University, Changchun, Jilin Province, China davidmedsci@sina.com.
Genet Mol Res ; 13(4): 8403-10, 2014 Oct 20.
Article em En | MEDLINE | ID: mdl-25366734
The insertion position of exogenous genes in plant genomes is usually identified by adapter ligation-mediated polymerase chain reaction (PCR), thermal asymmetric interlaced PCR, and restriction site extension PCR in transgenic plant research. However, these methods have various limitations, such as the complexity of designing primers and time-consuming and multiple-step procedures. The goal of this study was to establish an easier, more rapid, and more accurate method to clone flanking sequence using single-primer PCR in transgenic plants. Unknown flanking genome sequences in transgenic plants, including those in tobacco, soybean, rice, and maize, were cloned using the single-primer PCR method established in this study, with the Bar gene as the anchor gene. The primer 1 (P1), P2, and P3 PCRs obtained 4 sequences, and the completely correct flanking sequence of 508 bp that was obtained in the P3 PCR was verified by sequencing analysis. The single-primer PCR is more rapid and accurate than conventional methods, justifying its application widely in cloning flanking sequences in transgenic plants.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Plantas Geneticamente Modificadas / Clonagem Molecular Idioma: En Revista: Genet Mol Res Assunto da revista: BIOLOGIA MOLECULAR / GENETICA Ano de publicação: 2014 Tipo de documento: Article País de afiliação: China País de publicação: Brasil
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Plantas Geneticamente Modificadas / Clonagem Molecular Idioma: En Revista: Genet Mol Res Assunto da revista: BIOLOGIA MOLECULAR / GENETICA Ano de publicação: 2014 Tipo de documento: Article País de afiliação: China País de publicação: Brasil