关键词: 鹿茸, S100A4, 基因, 293T, 慢病毒载体, RNAi

Abstract: In this study,a number of sequences of small interfering RNA targeting S100A4 gene of sika deer (Cervus nippon)were synthesized and screened,among which two were confirmed by BLAST search to be suitable for the purpose. Oligo DNAs containing either sense or antisense strands were ligated into the lentiviral plasmids (pLVTHM)using T4 DNA ligase. Positive clones were identified based on the results of both PCR and sequencing. Each positive plasmid was cotransfected into 293T cells with the plasmids pCMV-dr 8. 91 and pMD2. G. Twenty-four hours after the co-transfection, green fluorescence of the 293 T cells could be observed under the inverted microscope. Therefore,we successfully constructed recombinant lentiviral system targeting sika deer S100A4 gene. This work would lay the foundation for revealing the regulatory mechanism of S100A4 underlying antler generation and regeneration.

Key words: Antler, 293 T, Lentiviral vector, RNAi, S100A4 gene