Construction of Tet-on inducible CXCR1 eukaryotic expression plasmid and identification of the expression character in NIH3T3 cells

Man Shi, Li bo Yao, Li feng Wang, Fu yang Li, Jin Su, Jian Zhang, Xia Li, Xin ping Liu

Research output: Contribution to journalArticlepeer-review

Abstract

AIM: To construct tetracycline (Tet)-controlled inducible vector CXCR1-pTREhyg, and then detect the expression character of CXCR1 under the regulation of Dox in NIH3T3 cells. METHODS: A full length cDNA of human CXCR1 was cloned from sample of fibroblast like synovium (FLS) in rheumatoid arthritis (RA) patient by RT-PCR and then sub-cloned into the pTREhyg plasmid after sequence analysis. CXCR1-pTREhyg and pTet-on was co-transfected with Lipofect2000 to NIH3T3 cells, and the expression of IL-8RA was detected by Western blot after given different concentration of Dox. RESULTS: Western blot showed that CXCR1 could be induced by Dox in NIH3T3 cells, and the phosphorylated Erk-1/2 level was significantly increased after IL-8 stimulation. CONCLUSION: Tet inducible recombinant vector of CXCR1-pTREhyg was successfully constructed, and it could be expressed in NIH3T3 cells. The stimulation of IL-8 obviously changed the activity of Erk-1/2 in the transfected NIH3T3 cells. This work has laid foundations for further study on the relationship between CXCR1 and RA disease.

Original languageEnglish (US)
Pages (from-to)280-282
Number of pages3
JournalXi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Volume22
Issue number3
StatePublished - May 2006

ASJC Scopus subject areas

  • Medicine(all)

Fingerprint

Dive into the research topics of 'Construction of Tet-on inducible CXCR1 eukaryotic expression plasmid and identification of the expression character in NIH3T3 cells'. Together they form a unique fingerprint.

Cite this