Human CD14 transgenic cell line.docVIP

 PAGE \* MERGEFORMAT 22 Human CD14 transgenic cell line Author: Ning Bo-tao, TANG Yong-min, Xu Yan, Chen Yanfei, Cao J Abstract In this study CD14 construct the eukaryotic expression plasmid to establish transgenic CD14-positive cell lines, for the establishment of acute monocytic leukemia (M5)-oriented treatment of animal models to provide research material. From normal human peripheral blood mononuclear cells extracted total RNA, a non-RNA enzyme DNA enzyme treatment, RT-PCR amplification of CD14 gene, TA cloned and sequenced with the GenBank sequence comparison of human CD14 gene to verify. By double digestion and in vitro ligation method to target gene was cloned into expression vector pcDNA 3.1 (); using Superfect transfection reagent to plasmid pcDNA 3.1 (+) / CD14 was transfected into C57BL / 6 mouse melanoma cell B16, by the G418 screened by flow cytometry the expression of CD14 protein in the initial screening out the CD14-positive cell lines B16/CD14. The results showed that: Sequencing and GenBank sequence comparison showed that the amplified gene sequences of the human CD14 is correct, and digestion results showed that the correct expression plasmid. Screened by flow cytometry the expression of two CD14-positive cell lines B16/CD14 (standard percentage of CD14-PE positive cells were 25.28%, 36.59%, 2F9-FITC for 25.59%, 36.32%). Conclusion: The human CD14 antigen expression in mouse cell lines B16/CD14, Weiren M5 animal model and its treatment-oriented research has laid a solid foundation. Keywords: transgenic cell line Establishment of Murine Cell Line Transfected with Human CD14 Gene Abstract This study was aimed to construct the CD14 eukaryotic expression vector, establish the transgeneic CD14 positive cell line in order to facilitate the establishment of a mouse model of antibody targeting therapy for human acute monocytic leukemia (AML-M5). Total RNA extracted from peripheral blood mononuclear cells was treated wi