Title: Construction of sense and antisense human heparanase and green fluorescent protein eukaryotic co-expressing vectors and their expression in lung cancer cell lines
Abstract: AIM: To construct sense and antisense human heparanase and enhanced green fluorescent protein eukaryotic expressing vectors and to transfect these vectors into lung cancer cell lines. METHODS: The human Heparanase cDNA fragment contained in the pcDNA3 Hpa vector was cloned into the enhanced green fluorescent protein eukaryotic expressing vector pIRES2 EGFP in cis direction or trans direction. The recombinant vectors were further identified by digestion of Bam H I and transfected to lung cancer cells by liposome method. After selection with G418, the transfected efficiency was observed under fluorescent inverted microscope. RESULTS: After digested by Bam H I, two fragments with the length of 5.3 and 1.7 kb, respectively, formed in sense fluorescent eukaryotic expressing vector (pIRES2 EGFP sHpa), while two other fragments with the length of 6.5 kb and 0.5 kb, respectively, formed in antisense fluorescent vector (pIRES2 EGFP aHpa). Electrophoretic results were completely identical with the theoretical calculation. Green fluorescence of the transfected cells was observed under fluorescent inverted microscope. CONCLUSION: Human heparanase sense and antisense fluorescent eukaryotic expressing vectors are successfully constructed and transfected to lung cancer cells.
Publication Year: 2004
Publication Date: 2004-01-01
Language: en
Type: article
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