West China Journal of Stomatology

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Prokaryotic Expression of Vascular Endothelial Growth Factor-C

PANJian*,WEN Yuming,HUAChengge,et al.   

  1. *Department ofOral andMaxillofacialSurgery,West China College ofStom- atology,Sichuan University,Chengdu610041,China
  • Received:2003-08-25 Revised:2003-08-25 Online:2003-08-20 Published:2003-08-20

Abstract:

Objective To evaluate whether the vascular endothelial growth factor (VEGF)-C cDNAwhich cloned from a patient with squamous cell carcinoma (SCC) of tongue can encode a functional protein or not.Methods RT-PCRwas employed to clone the functional VEGF-C fragment from the surgical specimen of a lingual SCC patient. Then it was subcloned into expres- sive plasmid vector pBKCMV, whichwas transfected intoE.colito examine its expression.Results Atruncated human VEGF- C cDNA fragment was amplified from the lingual SCC. The sequencing results of the fragment demonstrated that it had 99.6% similaritywith the reported human VEGF-C cDNA (representing the 559~1611 bp according the sequence of Genbank Entry X94216). Induced with IPTG, theE.coliXL1-Blue MRF′containing the recombinant pBK-VEGF-C expressed a 56 000 fusion protein, which can be recognized by polyclonic anti-humanVEGF-C antibody.Conclusion Afunctional fragmentVEGF-C cDNA was cloned from a lingual SCC. Itwill promote more intensive research on the function of VEGF-C and its relationshipwithmetas- tasis of oral SCC.

Key words: vascular endothelial growth factor (VEGF)-C, clone, prokaryotic expression