Construction and expression of human interleukin-13 and its mutant and subcellular location
- VernacularTitle:IL-13及其变异体真核表达载体的构建表达与亚细胞定位
- Author:
Yuwen WANG
;
Yongchen XU
;
Xiaofeng JIANG
- Publication Type:Journal Article
- Keywords:
Interleukin-13;
Mutant;
Green fluorescent protein;
Fusion protein;
Confocal microscopy
- From:
Chinese Journal of Immunology
1985;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct a eukaryotic expression vector for wild-type human Interleukin-13(whIL-13)and its mutant(mhIL-13) gene fused with enhanced green fluorescent protein(EGFP) and to analyze the expression and subcellular location of the fusion protein in COS-7 cells.Methods:The whole whIL-13 and mhIL-13 coding gene was amplified by RT-PCR. And then the expression vector for EGFP-whIL-13, EGFP-mhIL-13 fusion protein were constructed.The recombinant fusion gene were transfected into COS-7 cells.The expression and subcellular location of the fusion protein were detected by confocal microscopy.Results:The construction of expression vector were all corrected.In the positive cells, green fluorescent was found to be localized in cytoplasm,while empty in nucleus.Conclusion:The expression vectors for EGFP-whIL-13, EGFP-mhIL-13 fusion protein were constructed successfully and expressed in COS-7 cells. Fusion proteins were distributed in cytoplasm and no significant difference was observed between them.
