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    Construction and identification of eukaryotic expression vector expressing double shRNA sections targeting heparanase gene

¡¡¡¡¡¾Abstract¡¿ AIM: To construct and identify eukaryotic expression vector expressing double shRNA sections targeting heparanase gene (HpashRNA). METHODS:  Six pairs of oligonucleotides were designed and chemically synthesized. After randomly connecting an oligonucleotide and another one with 4-8 bases pairs interval, they were directionally inserted into plasmid pGenesil1 with respectively U6 promoter and termination code, the common green fluorescence protein (EGFP) gene and Neo gene. In this way, 3 vectors of pGenesil1HpashRNA containing 2 sections of HpashRNA were constructed and they were transfected into the ovarian cancer cell SKOV3. The rate of transfection was detected by flow cytometry and fluorescence microscope. The inhibition effectiveness of Hpa protein was analyzed by immunohistochemical staining. RESULTS:  The constructed eukaryotic expression vectors effectively suppressed the Hpa expression in transfected cells. The 3 vec

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