关键词: 血管内皮生长因子-C, 克隆, 原核表达

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