Isolation and Characterization of ABCB8 promoter gene of Danio rerio

Nur Razan Zafirah, Ramlee (2018) Isolation and Characterization of ABCB8 promoter gene of Danio rerio. [Final Year Project Report] (Unpublished)

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Abstract

ABCB8 gene is the ATP-Binding Cassette transporter protein in B subfamiy which are well known for multidrug resistance in cancer and multixenobiotics resistance against environmental pollution specifically in aquatic organisms. In human, ABCB8 gene has the role of mitochondiral ABC tranporter, iron homeostasis including protection towards oxidative stress. However, the research of the role of ABCB8 in Danio rerio has yet to discover. The aim of the research was to isolate and characterize on ABCB8 promoter gene in Danio rerio. In order to do isolation, total DNA extraction was done with TE buffer. A primer pair was designed using Primer Blast where forward and reversed primer that used in gradient PCR. The most suitable temperature to conduct gradient PCR along with primer pairs was 53.3°C with the size of 1384 bp. BLASTn was performed with sequencing result gave highly similarities according to the identity of 98% with E-value and gaps of 0.0. Restriction digestion was done with restriction enzyme of EcoRl and Sacl before ligated into pZsGreenl-l. Microinjection was done to further characterize ABCB8 gene in Danio rerio. Thus, the significant in ABCB8 gene in zebrafish could be identify and interaction that useful for ecotoxicological studies.

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