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[Construction of mutant myocilin plasmid and its expression in human trabecular meshwork cells]
Lina Wang1, Jian Ge, Bingqian Liu
1Key Laboratory of Ophthalmology of Ministry of Education and Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China.
Purpose:
To establish the mutant Myocilin gene plasmid in order to study the function of Myocilin.
Methods:
The mutant site was induced by site-directed mutagenesis. This plasmid were transfected into human trabecular meshwork cells (HTM cells) by cationic liposomes and the expression of Myocilin was examined by RT-PCR and Western blot analysis.
Results:
The mutant plasmid were correctly constructed. Myocilin was efficiently expressed in HTM cells transfected with this plasmid.
Conclusion:
The constructed eukaryote expression plasmid pcDNA-MYOC-P370L could express Myocilin in HTM cells in vitro.
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