title: Expression of recombinant carcinoembryonic-antigen-human-β-defensin-2 gene in colon cancer HCT116 cells.
abstract: The objective of this study was to construct a system driving high expression of human-β-denfensin-2 (HBD-2) system in eukaryotic cell. To construct recombinant retrovirus expression vector, CEA signal peptide-HBD-2 (mature peptide sequence) was cloned in the retrovirus expression vector PLNCX2. The retroviral vector was transfected into PT67 cells by DOTAP; after screening and amplifying single clone cell by G418, the virus particles were collected and infected to eukaryotic, colon cancer HCT116 cells. Furthermore, the HCT116 cells were also screened by using G418 and the resistance clones were obtained. Finally, the expression of HBD-2 was detected by Western blotting, which suggested a high level of expression of HBD-2 in HCT116 cells. In conclusion, the results indicate that high level of HBD-2 expression was obtained in HCT116 cells which will help in effective commercial production and purification of HBD-2 protein.
authors: Oncology Department, Anhui Provincial Tumor Hospital Affiliated to Anhui Medical University, Hefei, 230000, People's Republic of China.
authors: Yumei
authors: Y
authors: Zhang
authors: Jianping
authors: J
authors: Du
authors: Zhengtang
authors: Z
authors: Chen
mesh_terms: D002272:Carcinoembryonic Antigen / Q000235:genetics*; D003110:Colonic Neoplasms / Q000473:pathology*; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D045325:HCT116 Cells; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics; D017421:Sequence Analysis; D014162:Transfection; D023083:beta-Defensins / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D002272:Carcinoembryonic Antigen; C106641:DEFB4A protein, human; D011994:Recombinant Proteins; D023083:beta-Defensins