Objective    To establish the retrovirus expression vector containing glucose transporter-2 (GLUT-2) gene and its stable virus producing cell line，for the construction of new“artificial islet beta cells”.     Methods    Plasmid pCB7/GLUT2 was digested by EcoRⅠ/BamHⅠ and cloned to retroviral vector PLXSN for the construction of recombinant plasmid pL-GLUT2-SN

which was identified by enzyme digestion and sequencing

and then transfected to packaging cell line PA317.Titer of the recombinant virus was detected. Cell lines with a high titer were cloned and identified by PCR and RT-PCR.     Results    The retrovirus expression vec

tor pL-GLUT2-SN was established

in which the insertion site and reading frame of target gene were confirmed by enzyme digestion and sequence analysis. The highest titer of virus producing cell line PA317/GLUT2 was 7.1×10

5

CFU/mL. PCR and RT-PCR showed that the GLUT-2 gene was integrated into PA317/GLUT2 and stably expressed.     Conclusion    A retrovirus expression vector containing GLUT-2 gene and its stable virus producing cell line PA317/GLUT2 are successfully constructed

thus laying a foundation for the construction of “artificial islet beta cells”.