@article{ref1,
title="The construction of bicistronic eukaryotic vector carrying green fluorescent protein and hytk gene and its expression in bladder carcinoma",
journal="Chinese Journal of Pathophysiology",
year="1999",
author="Ye, Chuanzhong and Lin, Zhen and Chen, Shiping and Zhang, Fanglin and Pei, Xuetao and Li, Liang and Feng, Ka",
volume="",
number="12",
pages="-",
abstract="AIM: To facilitate the suicide gene delivery into neoplasm, a chimeric gene of HSV-tk and green fluorescent protein (gfp) was constructed. <br><br>METHODS: Molecular cloning technique was used to construct this kind of eukaryotic vector. The internal ribosome entry site (IRES) of encephalomyocarditis virus (EMCV), which could coordinate expression of two genes in a single vector, was optioned. By using liposome-mediated transfection, eukaryotic expression vector tgCMV/hytk-IRES-gfp was transfected into human bladder carcinoma cells EJ. <br><br>RESULTS: A bicistronic eukaryotic vector carrying gfp and hygromycin phosphotransferase-thymidine kinase fusion (hytk) gene was constructed. The results of PCR and microscopy detection show that the hytk-IRES-gfp gene was successfully transferred into EJ cells. There were no differences in the growth pattern or the morphology between EJ and EJ/hytk-GFP cells. In vitro experiments demonstrated dose- and time-dependent cell killing by transduction of the hytk-IRES-gfp gene followed by GCV treatment. The IC50 (the concentration required to elicit 50% growth inhibition) was 2.16 mg/L in treatment with GCV for 72 hours. <br><br>CONCLUSION: These results suggest that this new kind of eukaryotic vector could serves as a new tool and method for neoplasm gene therapy.<p /><p>Language: zh</p>",
language="zh",
issn="1000-4718",
doi="",
url="http://dx.doi.org/"
}