Molecular cloning and expression study of Grb 10 interacting GYF domain protein 1 (GIGYF 1) gene in zebrafish (Danio rerio) development

• Main Author: How, Kim Loong
• Format: Final Year Project Report
• Language: English; English
• Published: Universiti Malaysia Sarawak, (UNIMAS) 2011
• Subjects: QL Zoology
• Online Access: http://ir.unimas.my/id/eprint/5329/1/How.pdf; http://ir.unimas.my/id/eprint/5329/4/How%20Kim%20Loong.pdf; http://ir.unimas.my/id/eprint/5329/

Grb10 Interacting GYF domain (GIGYF) family genes consist of two members, namely GIGYF1 and GIGYF2 respectively. These two genes were initially identified through their interaction with Grb10, an adapter protein that binds activated Insulin-like Growth Factor I (IGF-1) and insulin receptors and may thus regulate IGF-I-stimulated growth. Recently, a study reported that GIGYF2 may play a causal role in familial Parkinson’s disease (PD). However, the function of GIGYF1 is largely unknown. Since both GIGYF1 and GIGYF2 are homologous proteins which share the common features, thus it would be interesting to find out the functional role of GIGYF1. Two GIGYF1 genes namely GIGYF1a and GIGYF1b were found in the zebrafish genome which consistent with the occurrence of genome duplication in zebrafish. Gene duplication event would enable further mutational study of GIGYF1 gene, especially in determining its relationship with PD which is stated in Duplication-Degenerative-Complementation (DDC) model. Total RNA was extracted from 10 different embryonic developmental stages (4cells, 3hpf, 5.25hpf, 11hpf, 18hpf, 24hpf, 36hpf, 48hpf, 3dpf, 5dpf) of zebrafish. The extracted RNA was then used to synthesize cDNA template and subjected to reverse transcription PCR (RT-PCR) analysis. The GIGYF1a and GIGYF1b gene were found to be expressed at all selected developmental stages and this might provide some hints to its functional role.