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| Construction of recombinant eukaryotic expression vector of human intestinal trefoil factor and its expression in CHO/dhfr- cells |
| ZHU Ronghe1, CHEN Huiyao2, ZHUANG Qiang2, JIANG Songfu2 |
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
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| Cite this article: |
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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.
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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.
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Received: 24 November 2015
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