title: Cloning of flanking sequence in transgenic plants by restriction site-anchored single-primer polymerase chain reaction.
abstract: Determining the insertion position of an exogenous gene in the target plant genome is one of the main issues in the transgenic plant field. This study introduced a simple, rapid, and accurate method to clone the flanking sequences of the transgenic bar gene as the anchoring gene in the transgenic maize genome using single-primer polymerase chain reaction (PCR). This method was based on the distribution of restriction sites in the maize genome and adopted the single-primer PCR method. Cloning the flanking sequences with the restriction site-anchored single-primer PCR simplified the experimental procedures by about 70% and reduced the experimental time by more than 80%. In conclusion, the restriction site-anchored single-primer PCR was a simple, rapid method to obtain the unknown flanking sequences in the transgenic plants.
authors.affiliation: Biotechnology Center of Jilin Agricultural University, Changchun, Jilin, China.
authors.forename: J
authors.initials: J
authors.lastname: Ma
authors.affiliation: Biotechnology Center of Jilin Agricultural University, Changchun, Jilin, China.
authors.forename: N N
authors.initials: NN
authors.lastname: Wang
authors.affiliation: Biotechnology Center of Jilin Agricultural University, Changchun, Jilin, China.
authors.forename: S
authors.initials: S
authors.lastname: Ren
authors.affiliation: Biotechnology Center of Jilin Agricultural University, Changchun, Jilin, China.
authors.forename: Y P
authors.initials: YP
authors.lastname: Fu
authors.affiliation: Biotechnology Center of Jilin Agricultural University, Changchun, Jilin, China.
authors.forename: S
authors.initials: S
authors.lastname: Lu
authors.affiliation: Biotechnology Center of Jilin Agricultural University, Changchun, Jilin, China.
authors.forename: Y P
authors.initials: YP
authors.lastname: Wang
authors.affiliation: Biotechnology Center of Jilin Agricultural University, Changchun, Jilin, China davidmedsci@sina.com.
authors.forename: P W
authors.initials: PW
authors.lastname: Wang
mesh_terms: D024506:5' Flanking Region / Q000235:genetics; D003001:Cloning, Molecular; D017931:DNA Primers; D018745:Genome, Plant; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D019076:Transgenes / Q000235:genetics*; D003313:Zea mays / Q000235:genetics*
chemical_list: D017931:DNA Primers