Molecular Analyses of Transgenic Plants

Kurniawan Rudi Trijatmiko; Felichi Mae Arines; Norman Oliva; Inez Hortense Slamet-Loedin; Ajay Kohli

doi:10.1007/978-1-4939-3289-4_15

Molecular Analyses of Transgenic Plants

Methods Mol Biol. 2016:1385:201-22. doi: 10.1007/978-1-4939-3289-4_15.

Authors

Kurniawan Rudi Trijatmiko¹, Felichi Mae Arines¹, Norman Oliva¹, Inez Hortense Slamet-Loedin¹, Ajay Kohli²

Affiliations

¹ Genetic Transformation Laboratory, Plant Breeding, Genetics and Biotechnology Division, International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines.
² Plant Molecular Biology Laboratory, Plant Breeding, Genetics and Biotechnology Division, International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines. a.kohli@irri.org.

PMID: 26614292
DOI: 10.1007/978-1-4939-3289-4_15

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.

Keywords: Chemiluminescent; Digoxigenin; Gene expression; SYBR Green; Southern blot; Transformation; Transgene; Western blot; qRT-PCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Blotting, Southern
Blotting, Western
Gene Expression
Oryza / genetics
Plants, Genetically Modified*
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
Transgenes / genetics*