[Construction of the eukaryotic expression vector pEGFP-claudin-1 and its expression in 293T cells].

AIM

To construct the eukaryotic expression vector pEGFP-claudin-1 and to make it express expressed in 293T cells.

METHODS

Claudin-1 ORF was amplified by reverse transcription polymerase chain reaction (RT-PCR). The eukaryotic expression vector pEGFP-claudin-1 was constructed by introducing claudin-1 DNA fragment into the sites of Xho I and BamH I of pEGFP-C3 vector. The plasmid was transfected into 293T cells with lipofectamine. The expressed EGFP was observed by fluorescent microscope. The EGFP-claudin-1 was detected by Western blot.

RESULTS

The eukaryotic expression vector pEGFP-claudin-1 was constructed successfully. The expression of EGFP-claudin-1 was observed in the membrane of transfected cells by fluorescent microscope and a 49 kD protein was detected by Western blot.

CONCLUSION

The recombinant expression vector pEGFP-claudin-1 has been successfully constructed and claudin-1 gene can be expressed in 293T cells. This study lays a foundation for further research into the function of claudin-1.