[Construction of eukaryotic expression vector of CD80-IgG1 Fc fragment fusion protein and its expression in CHO cells].
OBJECTIVE
To construct an eukaryotic expression vector of CD80-IgG1 Fc, and to express the fusion protein in CHO cells.


METHODS
The gene encoding the CD80-IgG1 Fc fusion protein were constructed in eukaryotic expression vector pcDNA3.1(+) by means of T-A cloning and subcloning techniques, then was transfected into CHO cells for stable expression. The expression of the fusion protein was detected by Western blot and ELISA.


RESULTS
DNA sequencing and restriction endonuclease digestion analysis indicated that the eukaryotic expression vector CD80-IgG1 Fc/pcDNA3.1(+) was successfully constructed. After the recombinant plasmid was transfected into CHO cells, the stable expression of the fusion protein was demonstrated by Western blot and ELISA.


CONCLUSIONS
The eukaryotic expression vector of CD80-IgG1 Fc/pcDNA3.1(+) was successfully constructed and stably expressed in CHO cells, providing basis for anti-tumor study.
Li
He-jun
HJ
Institute of Clinical Immunology, Union Hospital Affiliated to Fujian Medical University, Fuzhou 350001, China.
Li
Pin
P
Zhou
Xiao-ling
XL
Zheng
Xiang-xiong
XX
D000818:Animals; D018122:B7-1 Antigen / Q000235:genetics*; D015153:Blotting, Western; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics*; D007141:Immunoglobulin Fc Fragments / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism*
D018122:B7-1 Antigen; D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins