1.Department of Pediatric, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015; 2.Department of Hematology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015
ZHU Ronghe,CHEN Huiyao,ZHUANG Qiang, et al. Construction of recombinant eukaryotic expression vector of human intestinal trefoil factor and its expression in CHO/dhfr- cells[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2016, 46(8): 561-565.
Abstract：Objective: To construct eukaryotic expressing vector of human intestinal trefoil factor and express hITF protein in dihydrofolate reductase-deficient Chineses hamster ovary cells (CHO/dhfr-). Methods: The full length cDNA fragment was amplified by overlap extension PCR, then inserted into eukaryotic expression vector pIRES/dhfr. The recombinant vector was transfected into CHO/dhfr- cells by LipofectamineTM2000. The stable transfected CHO/dhfr- cell line was then established by screening cultures with G418. RT-PCR and ELISA were used to detect the expression of the protein. Results: The eukaryotic expression vector was constructed successfully, stable transfected CHO cell line was established, and hITF protein was expressed successfully. Conclusion: The recombinant plasmid pIRES/hITF/dhfr is successfully constructed and it expresses the protein in the supernatant of the CHO/dhfr- cells, which is the basis for pre-clinical study.
