Construction and identification of PRDX3 eukaryotic expression vector
10.7644/j.issn.1674-9960.2016.10.006
- VernacularTitle:PRDX3真核表达载体构建及亚细胞定位
- Author:
Chunxia ZHANG
;
Long CHENG
;
Hongxu MAI
;
Lin WANG
;
Juhui ZHANG
;
Enqun WANG
;
Qinong YE
- Publication Type:Journal Article
- Keywords:
peroxiredoxin3;
clone;
eukaryotic expression;
subcellular localization
- From:
Military Medical Sciences
2016;40(10):801-804,808
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the eukaryotic expression vector of PRDX3 labeled with FLAG tag and to study its localization in human tongue cancer cell line SCC15.Methods PRDX3 gene was obtained from the breast library by PCR and cloned into PCDH vector to construct PCDH-FLAG-PRDX3.The plasmid was transiently transfected into 293T cells and the expression was detected by Western blot.Subcellular localization was detected by cellular immunofluorescence.Results The result of double digestion and sequencing showed that PCDH-FLAG-PRDX3 eukaryotic expression vector was constructed.The expression of FLAG-PRDX3 in human 293T cells was positively confirmed by Western blotting.In human tongue cancer cell line SCC15, the result of cellular immunofluorescence showed FLAG-PRDX3 was located in the cytoplasm rather than in the nucleus.Conclusion PRDX3 eukaryotic expression vector labeled with FLAG tag is constructed successfully, which is located in cytoplasm in human SCC15 cells.Construction and identification of PRDX3 could shed light on the function and mechanism of PRDX3 in tongue cancer.
