Prokaryotic expression of vascular endothelial growth factor-C.
- Author:
Jian PAN
1
;
Yuming WEN
;
Chengge HUA
Author Information
- Publication Type:Journal Article
- MeSH: Carcinoma, Squamous Cell; genetics; metabolism; Cloning, Molecular; DNA, Complementary; biosynthesis; genetics; Escherichia coli; genetics; Humans; Lymphatic Metastasis; Prokaryotic Cells; metabolism; Tongue Neoplasms; genetics; metabolism; Transfection; Vascular Endothelial Growth Factor C; biosynthesis; genetics
- From: West China Journal of Stomatology 2003;21(4):321-323
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo evaluate whether the vascular endothelial growth factor (VEGF)-C cDNA which cloned from a patient with squamous cell carcinoma (SCC) of tongue can encode a functional protein or not.
METHODSRT-PCR was employed to clone the functional VEGF-C fragment from the surgical specimen of a lingual SCC patient. Then it was subcloned into expressive plasmid vector pBKCMV, which was transfected into E. coli to examine its expression.
RESULTSA truncated human VEGF-C cDNA fragment was amplified from the lingual SCC. The sequencing results of the fragment demonstrated that it had 99.6% similarity with the reported human VEGF-C cDNA (representing the 559-1611 bp according the sequence of Genbank Entry X94216). Induced with IPTG, the E. coli XLI-Blue MRF' containing the recombinant pBK-VEGF-C expressed a 56,000 fusion protein, which can be recognized by polyclonic anti-human VEGF-C antibody.
CONCLUSIONA functional fragment VEGF-C cDNA was cloned from a lingual SCC. It will promote more intensive research on the function of VEGF-C and its relationship with metastasis of oral SCC.