Identification of differentially expressed genes during somatic embryogenesis of Axonopus compressus by restriction fragment differential display-coupled FSD

• Main Authors: See, Pao Theen, Abdul Rahman, Siti Suhaila, Chin, Chiew Foan, Kulaveerasingam, Harikrishna
• Format: Article
• Language: English
• Published: Malaysian Society for Molecular Biology and Biotechnology 2003
• Online Access: http://psasir.upm.edu.my/id/eprint/58524/1/Identification%20of%20differentially%20expressed%20genes%20during%20somatic%20embryogenesis%20of%20Axonopus%20compressus%20by%20restriction%20fragment%20differential%20display-coupled%20FSD.pdf; http://psasir.upm.edu.my/id/eprint/58524/; http://www.msmbb.org.my/apjmbb/html112/112.htm
• Institution: Universiti Putra Malaysia

We present an approach to profile the expression of gene family members during somatic embryogenesis of Axonopus compressus through the modification of a recently developed RNA fingerprinting method. The protocol combines restriction fragment differential display technology with primers directed to a gene family specific domain signature on mRNA and hence is termed ‘RFDD-coupled FSD’. By using this method, two differentially expressed mRNAs, AC1 and AC2, containing the chromo-like domain and the serine/threonine protein kinase signatures, respectively, were isolated. Sequence analysis revealed that AC1encoded a MIP (plasma membrane protein) meanwhile AC2 shared low homology to other serine/threonine protein kinases. The abundance of these two transcripts appeared to be down-regulated in the embryogenic and non-viable embryogenic tissues when examined on Northern and semi-quantitative RT-PCR analyses, suggesting possible developmental arrest. This preliminary differential expression examination has somewhat provided a first insight into the molecular events underlying A. compressus somatic embryo development. Thus, RFDD-coupled FSD has been shown to be an efficient procedure for identifying differential regulated transcripts that correspond to their defined gene family domain signature sequences.