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Construction of eukaryotic vector of human neuronal pentraxin 2 and establishment of stable transfected PANCI cell line

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Author:
No author available
Journal Title:
CHINESE JOURNAL OF PANCREATOLOGY
Issue:
1
DOI:
10.3760/cma.j.issn.1674-1935.2010.01.012
Key Word:
胰腺肿瘤;NPTX2;转染;细胞系;Pancreatic neoplasms;NPTX2;Transfection;Cell line

Abstract: Objective To construct eukaryotie vector of human neronal pentraxin 2 (NPTX2),and obtain the sstable transfected PANC1 cell lines.Metods The full-length cDNA of NPTX2 was diget EcoRl and Notl,and was cloned into plasmid pcDNA3,1,which were confirmed by sequencing,then the PcDNA3,1(+)and pcDNA3,1(+)-NPTX2 vectors were transfected into PANC1 cells by LipofectamineTM 2000,The stable transfected PANC1 cell lines were selected by the abiliy of resistanc to G418.The NPTX2 mRNA expression of PANC1 in the selected clones was detected by real-time quantitative PCR.Results The eukaryotic vector pcDAN3,1(+)-NPTX2 was constructed successfully,stable transfected PANC1 cell line was established and confirmed by real-time quantitative PCR.Conclusions The construction of eukaryotic vector targeting NPTX2 and the established sstable transfected PANC1 cell line provided a solid experimental foundation for further studies on the function of NPTX2 in pancreatic cancer cells.

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