Introduction of the Serine Green Fluorescent Protein (sGFP) Gene into Pyricularia grisea Race dc4 Isolated from Digitaria ciliaris using Agrobacterium tumefaciens-mediated Genetic Transformation

Stephanie Stephanie, Utut Widyastuti, Suryo Wiyono

Abstract


Gene serin Green Fluorescent Protein (sGFP) has been used to monitor gene expression specific tagged proteins that has implication for fungal cell study. This research aimed to introduce sGFP gene into genome of P. grisea dc4 from D. ciliaris using A. tumefaciens. Plasmid sGFP was introduced into A. tumefaciens by triparental mating method (TPM). Genetic transformation was performed by co-cultivating spore P. grisea dc4 with A. tumefaciens LBA4404–pCAMB-sGFP. Pyricularia grisea dc4 transformant was selected by using selection medium that contains 300 µg/ml hygromycin. The integration of sGFP gene into genome was confirmed by PCR using  sGFP’s spesific primer pair, sGFP-Nos terminator primer pair and β-Tubulin primer pair as internal control. Expression of sGFP from P. grisea dc4 transformant were detected with blue light fluorescent microscope.


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DOI: http://dx.doi.org/10.11598/btb.2015.22.1.329

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