Far red fluorescent protein HcRed prokaryotic and eukaryotic expression and identification of.docVIP

 PAGE \* MERGEFORMAT 13 Far red fluorescent protein HcRed prokaryotic and eukaryotic expression and identification of Study: Yuan Shi Yan Xue Heng Yang Zhiqiang Zhao Yinxia Wang Kai Wang Shengjun Shaoqi Xiang Jiao Zhijun Xu Wen Elite of River [Abstract] Objective: gene were constructed to carry HcRed PET28a (+) prokaryotic and eukaryotic expression vector PIRES, respectively, in E. coli and mouse dendritic cells (DC2.4) in the expression, identification, and for the subsequent gene protein transfection provided. Methods: According to the gene pool HcRed1 gene sequence HcRed CDS full length primers containing HcRed plasmid as a template by PCR to obtain the target gene, and were connected to the PET28a (+) prokaryotic vector and PIRES eukaryotic vector. by PCR, restriction analysis, the original nuclear vector was transformed into Escherichia coli, induced by IPTG; eukaryotic expression vector using Lipofectamine DC2.4, G418 selected clones, the application of RT PCR and flow cytometry for detection. Results: amplified 687 bp of HcRed CDS sequence, prokaryotic and eukaryotic expression vector PET28a (+) / HcRed and PIRES / HcRed, respectively, in Escherichia coli and DC2.4 cell lines successfully expressed. CONCLUSION: The constructed HcRed prokaryotic, eukaryotic expression vector, respectively, strains and cell lines in the project was successfully expressed as a fusion protein gene and the follow-up screening, tracing and in vivo observations laid the foundation. [Keywords:] far red fluorescent protein HcRed; 2.4 Dendritic cells prokaryotic expression; [Abstract] Objective: In order to use the far red fluorescent protein HcRed, we amplified HcRed gene and cloned into PET28a (+) and PIRES plasmids. Then, the HcRed was expressed and identified in E.coli and Dendritic cell line, which provided the basis for further research.Methods: The primers were designed with HcRed gene in GenBank. Using standard PCR and cloning techniques, the HcRed gene was amp