People notch1shRNA lentiviral expression vector and identification of.doc

 PAGE \* MERGEFORMAT 18 People notch1shRNA lentiviral expression vector and identification of Of: Zhang Yan, NANJING RADIO AND TELEVISION, LIN Ling, ZHENG Zhi MATRIX [Abstract] Objective To construct notch1 shRNA Lentiviral expression vector for the Notch signaling pathway related basis for the study. Methods The mRNA sequence of human notch1 gene and DNA synthesis of two complementary single strand oligonucleotides, formed after the annealing connected pENTR/U6 double-stranded plasmid with pLenti6/BLOCK iT DEST by LR recombination vector was constructed notch1 shRNA lentiviral vector, liposome into 293FT cells, packaged into lentivirus. with the slow virus infection of human neural neuroblastoma line SH SY5Y cells, RT PCR and Western blot, cell notch1 gene expression. Results successfully connected to the target sequence vector and packaged into a high titer lentivirus, cells were detected mRNA and protein showed that the constructed notch1 Lentiviral shRNA expression vector LR ND A can inhibit the SH SY5Y cells notch1 gene expression. Conclusion constructed notch1 shRNA lentiviral vector. [Keywords:] RNA gene expression in lentivirus vector signal transduction of genetic Notch signaling pathway and human and animal embryonic development, cell proliferation and differentiation and is closely related to tumorigenesis, Notch receptor is a class of 2753 amino acid transmembrane receptor, the homologous molecules, including Notch l ~ 4. Notch by ligand DSL (Delta Serrate Lag 2) activation of protein binding family, you can switch the cell cycle of conversion, regulation of apoptosis, survival and differentiation fate choice, as an essential component of Notch signaling pathway also Notch1 receptor become an important target of study [1 2]. RNA interference (RNAi) technology can be effective, specifically reduced the expression of target genes is a simple and rapid means of gene function [3]. lentiviral expression vector transfected into mammalian cells