Construction of murine CCL21 eukaryotic expression plasmid and chemotactic function of its products.
- Author:
Ting GONG
1
;
Hong-li LI
;
Yi BA
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Base Sequence; Chemokine CCL21; biosynthesis; genetics; Chemotaxis, Leukocyte; Cloning, Molecular; DNA, Complementary; genetics; Dendritic Cells; drug effects; immunology; Genetic Vectors; genetics; Mice; Mice, Inbred C57BL; Molecular Sequence Data; Plasmids; genetics; Recombinant Proteins; biosynthesis; genetics; immunology; Stomach Neoplasms; metabolism; pathology; Transfection; Tumor Cells, Cultured
- From: Journal of Zhejiang University. Medical sciences 2009;38(2):125-129
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the murine CCL21 eukaryotic expression plasmid, and to investigate the chemotactic function of its products.
METHODSMurine CCL21 cDNA was amplified by RT-PCR from murine total RNA, and was inserted into eukaryotic expression plasmid pcDNA3.1 after confirmation of sequencing. The recombinant CCL21 plasmid was transferred into mouse forestomach carcinoma (MFC) cells and the chemotactic function of expressed products was detected by chemotaxis assay.
RESULTGene sequencing, gel electrophoresis of PCR products and restrictive digestion proved the successful construction of CCL21, and its expression was confirmed by Western Blot. The transfected tumor cells had a significant chemotactic function to DC.
CONCLUSIONThe recombinant murine CCL21 eukaryotic expression plasmid has been successfully constructed, and its expression products in tumor cells have a marked chemotactic function to DC.
