Isolation and Characterization of ABCB5 Gene Promoter Region from Zebrafish
ABCB5 (ATP-binding cassette, subfamily B, member 5) protein consists of P-glycoprotein that affects the levels of plasma and intracellular drugs concentration that control therapeutic response. This study aims to isolate and characterize the promoter region of the ABCB5 gene from zebrafish (Danio...
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| Main Author: | |
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| Format: | Final Year Project Report |
| Language: | English |
| Published: |
Universiti Malaysia Sarawak (UNIMAS)
2018
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| Online Access: | http://ir.unimas.my/id/eprint/35384/1/Auni%20Haziqah%20%20ft.pdf http://ir.unimas.my/id/eprint/35384/ |
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| Summary: | ABCB5 (ATP-binding cassette, subfamily B, member 5) protein consists of P-glycoprotein
that affects the levels of plasma and intracellular drugs concentration that control
therapeutic response. This study aims to isolate and characterize the promoter region of the
ABCB5 gene from zebrafish (Danio rerio). Initially, DNA extraction was performed on
zebrafish by using lysis buffer. Then, the primer for the ABCB5 promoter was designed
and the gradient PCR was done with the optimum temperature 53.5°C to produce single
amplicon with the approximate size of 1,500 bp. The amplicon was cloned into pGEM-T
Easy Vector and transformed into Escherichia coli XLl Blue Competent cells with the
transformation efficiency of 0.05x10° transform ants per µg. Colony PCR was conducted to
determine the positive colony with promoter insert. Next, plasmid miniprep was conducted
before sending the plasmid for sequencing. However, the BLASTn results showed that the
sequence cloned is not ABCB5 promoter region from zebrafish. Hence, it is further
suggested that cloning should be continued in order to determine the transcription factor
which regulates ABCB5 in order for us to further und |
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