Molecular cloning and expression studies of growth factor receptor bound protein 14 (Grb14) gene in zebrafish development

Chin, Louis Vui Ngian (2011) Molecular cloning and expression studies of growth factor receptor bound protein 14 (Grb14) gene in zebrafish development. [Final Year Project Report] (Unpublished)

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Abstract

Grbl4 was the most recently identified members of Grb7 superfamily of adaptor protein, which participate in the functionality of multiple signal transduction pathways under the control of a variety of activated tyrosine kinase receptors (RTK). To date, the functional roles of Grbl4 adaptors is not fully understood. At the present time, a role for this GrbI4-RTK interaction was only established in the regulatory effect on RTK signaling especially in insulin receptor signaling inhibition as well as cell migration and tumor progression. In respect that most of the current knowledge and understanding of Grb14 expression come from murine studies, therefore, in this project the expression studies of Grb14 will be determined from 10 different developmental stages in Zebrafish (Danio rerio) which comprise of 4-cells, 3hpf, Shpf, Ilhpf, 18hpf, 24hpf, 36hpf, 48hpf, 3dpf and Sdpf. From Reverse Transcription (RT -PCR) , the expression degree from each respective stage will allowed us to comprehend when or which developmental stage that Grb14 started to be expressed and how strong the expression degree in distinct developmental stage. Based on the RT-PCR semi-quantitative analysis result of gene expression in this study, Grb14 was expressed in all the embryonic developmental stages of zebrafish with dissimilitude in expression degree. Beside that, direct sequencing result from BLAST showed that the nucleotide sequences exhibit 99% of homology similarity from Grbl4 NCBI database. Later on, Grb14 which had been successfully cloned can be used in assisting in future functional studies of Orb14 using in situ hybridisation followed by gene known down approach .

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