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Construction and identification of recombinant plasmid pUIS3-BLC+

文献类型： 外文期刊

作者： Lin, Yingzi 1 ; Bao, Shixiang 2 ; Tan, Guanghong 1 ; Huang, Fengying 1 ; Huang, Huiqin 2 ;

作者机构： 1.Hainan Med Univ, Hainan Prov Key Lab Trop Med, Haikou 571101, Peoples R China

2.Hainan Univ, Sch Agr, Haikou 571101, Peoples R China

3.Chinese Acad Trop Agr Sci, Biol Technol Inst, Haikou 571101, Peoples R China

关键词： BLC;Plasmodium berghei;Transgene;Knockout

期刊名称：ASIAN PACIFIC JOURNAL OF TROPICAL MEDICINE （ 影响因子：1.226； 五年影响因子：2.285 ）

ISSN： 1995-7645

年卷期： 2010 年 3 卷 10 期

页码：

收录情况： SCI

摘要： Objective: To construct and identify recombinant plasmid pUIS3-BLC+. Methods: The cDNA of B-lymphocyte chemoattractant (BLC) was amplified from the total RNA of spleen tissues by PCR method, and were inserted into plasmid of Plasmodium berghei with UIS3 knockout by digestion of restrictive endonuclease and T7 ligation. The recombinant plasmids were screened, and then underwent restriction enzymatic digestion and DNA sequencing. Then the confirmed plasmid was further transfected into COS-1 cells by lipofectamine and the BLC expression was tested by RT-PCR and Western blotting. Results: The cDNA of BLC gene was correctively amplified by RT-PCR and the recombinant plasmid pUIS3-BLC+ was constructed successfully, which was confirmed by restriction enzymatic digestion and DNA sequencing. RT-PCR and Western blotting also showed the BLC gene expression in COS-1 cells. Conclusions: The recombinant plasmid pUIS3-BLC+ has BLC expression in COS-1 cells, and is useful for further study on BLC transgene and UIS3 gene knockout in Plasmodium berghei.

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