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Published on: February 9, 2015
[Construction and expression of human IL-8 sense or antisense eukaryotic expression vectors]
Xiu-long Niu1, Yue Wang, Ye Qu
1Department of Immunology, Medical College of Chinese People's Armed Police Forces, Tianjin 300162, China. nxlwi_ren@126.com
Aim:
To construct the sense or antisense IL-8 eukaryotic expression vectors.
Methods:
Sense or antisense IL-8 full length gene were amplified by RT-PCR and cloned into eukaryotic expression vector pcDNA3.1(+). After the identification by PCR, restriction endonuclease digestion and the nucleotide sequencing, the recombinant vectors were transfected into human ovarian carcinoma A2780 and SKOV3 cell lines transiently by lipofectamine mediation. The expression of IL-8 gene and protein were detected by RT-PCR and ELISA.
Results:
The sense or antisense IL-8 eukaryotic expression vectors were constructed and verified. The expression of IL-8 gene and protein in A2780 cells transfected with pcDNA3.1(+)-ssIL-8 were increased, whereas the expression of IL-8 protein in SKOV3 cells transfected with pcDNA3.1(+)-asIL-8 was decreased.
Conclusion:
The eukaryotic expression vectors pcDNA3.1(+)-ssIL-8 or pcDNA3.1(+)-asIL-8 have been constructed successfully, which lays a base for further study on roles of IL-8 in ovarian cancer and other tumors.
