环状RNA mmu-circ-Pax3.1表达载体的构建及初步验证

doi:10.16431/j.cnki.1671-7236.2017.10.006

Abstract

Abstract:

This study was aimed to investigate the feasibility of expressing mmu-circ-Pax3.1 by circular RNA expressed vector in vitro. The exists of mmu-circ-Pax3.1 was confirmed by reverse PCR and sequenced, and constructed pcDNA3.1(+) CircRNA-mmu-Pax3.1 through cloning the corresponding linear sequence into pcDNA3.1(+) CircRNA Mini Vector. The recombinant vector was identified by PCR, enzyme digestion and sequencing methods.The recombinant plasmid was transfected into 293 cells by Lipofectamine 2000 transfection reagent, transfected cells were observed under the inverted fluorescence microscope. Finally,the expression of mmu-circ-Pax3.1 was detected by RT-PCR in normal cells group, empty vector group and experimental group. The mmu-circ-Pax3.1 expression vector was successfully constructed, and the vector could make mmu-circ-Pax3.1 efficient transcription into 293 cells. The circular RNA expression vector constructed in this experiment by genetic engineering technology was transfected into 293 cells by liposome method, and transcribed mmu-circ-Pax3.1 efficiently and laid a foundation for further study for the function of mmu-circ-Pax3.1.

Key words: circular RNA; vector construction; 293 cells; transfection

CLC Number:

CAO Yang, YOU Shuang, YAO Yang, LI Cun-yuan, CHEN Chuang-fu, NI Wei, HU Sheng-wei. Construction and Preliminary Verification of Expression Vector of Circular RNA mmu-circ-Pax3.1[J]. , 2017, 44(10): 2865-2870.

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Construction and Preliminary Verification of Expression Vector of Circular RNA mmu-circ-Pax3.1

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