pcEgr-IFNγ重组质粒的构建与体外X射线诱导表达特性

doi:10.3969/j.issn.1004-616x.2003.01.001

Abstract

Abstract: Purpose:Cloning IFNγcDNA and constructing pcEgr-IFNγto study its expression intransfected B16 cells induced by X-ray irradiation indifferent doses,and to study the time course of the expression. Methods:IFNγcDNA was cloned by PCR and inserted into pcDNA3.1.CMV promoter of pcDNA3.1 was replaced by Egr-1 promoter to construct pcEgr-IFNγplasmid.The plasmids were transfected into B16 cells.The expression of IFNγof transfected B16 cells with X-ray irradiation indifferent doses and expression time course were detected by ELISA.Results:PCR product was sequenced,there sultindicated the sequence was correct.There combined plasmid was detected by electrophoresis.After X-ray irradiation indifferent doses,IFNγexpression of transfected B16 cells was 77.73 ～ 94.60pg / ml,significantly higher than the control group(P<0.05 ～ P<0.01).Six hours after 2 Gy X-ray irradiation,the expression was the highest(90.00pg / ml),significantly higher than the control group(P<0.001).Conclusion:X-ray caninduce the recombine plasmid expressin B16 cells.The detection of doses and time course provides an experimental basis for invivo study infuture.

Key words: IFNγexpression, PCR, Construction of plasmid, X-ray irradiation, Time course, Dose effect

CLC Number:

R146

WU Cong-mei , LI Xiu-yi . STUDY ON CONSTRUCTION OF pcEgr-IFNγ AND EXPRESSION PROPERTY OF X-RAYINDUCTION IN VITRO[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2003, 15(1): 1-4.

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STUDY ON CONSTRUCTION OF pcEgr-IFNγ AND EXPRESSION PROPERTY OF X-RAYINDUCTION IN VITRO

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