title: Molecular Analyses of Transgenic Plants.
abstract: One of the major challenges in plant molecular biology is to generate transgenic plants that express transgenes stably over generations. Here, we describe some routine methods to study transgene locus structure and to analyze transgene expression in plants: Southern hybridization using DIG chemiluminescent technology for characterization of transgenic locus, SYBR Green-based real-time RT-PCR to measure transgene transcript level, and protein immunoblot analysis to evaluate accumulation and stability of transgenic protein product in the target tissue.
authors.affiliation: Genetic Transformation Laboratory, Plant Breeding, Genetics and Biotechnology Division, International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines.
authors.forename: Kurniawan Rudi
authors.initials: KR
authors.lastname: Trijatmiko
authors.affiliation: Genetic Transformation Laboratory, Plant Breeding, Genetics and Biotechnology Division, International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines.
authors.forename: Felichi Mae
authors.initials: FM
authors.lastname: Arines
authors.affiliation: Genetic Transformation Laboratory, Plant Breeding, Genetics and Biotechnology Division, International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines.
authors.forename: Norman
authors.initials: N
authors.lastname: Oliva
authors.affiliation: Genetic Transformation Laboratory, Plant Breeding, Genetics and Biotechnology Division, International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines.
authors.forename: Inez Hortense
authors.initials: IH
authors.lastname: Slamet-Loedin
authors.affiliation: Plant Molecular Biology Laboratory, Plant Breeding, Genetics and Biotechnology Division, International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines. a.kohli@irri.org.
authors.forename: Ajay
authors.initials: A
authors.lastname: Kohli
mesh_terms: D015139:Blotting, Southern; D015153:Blotting, Western; D015870:Gene Expression; D012275:Oryza / Q000235:genetics; D030821:Plants, Genetically Modified; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes / Q000235:genetics*
keywords: Chemiluminescent; Digoxigenin; Gene expression; SYBR Green; Southern blot; Transformation; Transgene; Western blot; qRT-PCR