Construction of an inducible green fluorescent protein (GFP) gene in a binary vector
Green fluorescent protein (GFP) gene is synthesized by jellyfish Aequorea victoria and it is widely used as reporter gene in molecular biology studies. The discovery of this reporter gene contributes to the development in the gene expression, cell development and signal transduction studies. It al...
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| Format: | Final Year Project Report / IMRAD |
| Language: | English |
| Published: |
Universiti Malaysia Sarawak, UNIMAS
2013
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| Online Access: | http://ir.unimas.my/id/eprint/7490/ http://ir.unimas.my/id/eprint/7490/14/Nurul%20Liana%20Mohd%20Khalid%20ft.pdf |
| Summary: | Green fluorescent protein (GFP) gene is synthesized by jellyfish Aequorea victoria and it is widely used as reporter
gene in molecular biology studies. The discovery of this reporter gene contributes to the development in the gene
expression, cell development and signal transduction studies. It also leads to the discovery of novel characteristics in
a particular organism. The construction of inducible synthetic GFP into a binary vector was carried out by using the
restriction enzyme digestion of the DNA fragments and followed by the ligation of the DNA fragments by DNA
ligase. Both mechanisms were carried out in vitro. The binary vector was used in this research because it facilitates
the transfer of the gene into a target organism via Agrobacterium- mediated transformation. Meanwhile, for the
verification steps, the Polymerase Chain Reaction (PCR) was used to identify the integration of synthetic GFP gene
in the binary vector. There were many Escherichia coli XL1 Blue colonies grew on the Luria Agar (LA) media
containing antibiotic ampicillin. However, the PCR cannot verify the presence of sGFP gene in the pAGS3/sGFP
plasmids harbored by the E. coli XL1 Blue colonies. The result indicated that there was an absence of pAGS3/sGFP
plasmid in the transformed E. coli XL1 Blue. |
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