Identification of AP1 cis-element and transcriptional effect on cytokeratin 13 gene expression.
- Author:
Gongbiao LIN
1
;
Jianyun XIAO
;
Suping ZHAO
;
Yongquan TIAN
;
Chenglong WANG
;
Yuanzheng QIU
Author Information
- Publication Type:Journal Article
- MeSH: 5' Flanking Region; genetics; Base Sequence; Binding Sites; genetics; Binding, Competitive; Chloramphenicol O-Acetyltransferase; genetics; metabolism; DNA; genetics; metabolism; Electrophoretic Mobility Shift Assay; Gene Expression Regulation; HeLa Cells; Humans; Keratins; genetics; Mutation; Recombinant Fusion Proteins; genetics; metabolism; Transcription Factor AP-1; metabolism; Transcription, Genetic; genetics; Transfection
- From: Chinese Journal of Medical Genetics 2002;19(6):479-483
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo identify the type of CTGAATCA from -nt.199 to -nt.192 of the cytokeratin 13(CK13) gene 5' flanking region and determine its transcriptional effect on CK13 gene expression.
METHODSThe CAT systems were used to assess the effects of different motifs of CK13 gene 5' flanking region on transcription. The clones of pCAT-enhancer with the total length, -nt.207 to +nt.63 and the same length of -nt.207 to +nt.63, but the T, G of -nt.198, -nt.197 being changed to A, T of the CK13 gene 5' flanking region, were constructed and transferred to HeLa cells with the help of lipofectin. Then work was done to detect the instant CAT expression of different clones and evaluate the effects of CTGAATCA of the 5' flanking region on CK13 gene expression. The type of the cis-element of CTGAATCA was identified with electrophoretic mobility shift assay (EMSA) and competition-EMSA.
RESULTSCTGAATCA in the CK13 gene 5' flanking region is an AP1 cis-element by EMSA and competition-EMSA, it promotes CK13 gene expression.
CONCLUSIONCTGAATCA from -nt.199 to nt.192 of the CK13 gene 5' flanking region is an AP1 reaction element, not a cAMP reaction element. It promotes transcriptional activity of CK13 gene 5' flanking region.
