吉林农业大学学报Issue(1)：116-121,6.DOI:10.13327/j.jjlau.2014.1719
利用慢病毒表达载体干扰梅花鹿角柄骨膜细胞P21基因
RNA Interference Targeting P21 Gene of Pedicle Periosteum Cells from Sika Deer by shRNA Lentivirus
摘要
Abstract
P21 gene of the pedicle periosteal cells of sika deer was interfered using RNAi in lentiviral vector system .The results showed that:(1 )Two sequences of small interfering RNAs,targeting P21 gene of sika deer,were successfully reassembled into the lentiviral plasmids(Plvthm).Positive clones were identified based on the results of both PCR and sequencing .Recombinant lentivirus was acquired by each positive plasmid co-transfecting into 293 T cells with the plasmids pMD2 .G and pCMV-dr8.9;(2)Recombinant lentivirus was successfully interfered into the PP cells,and the GFP positive cell pro-portion obtained by flow cytometry (FCM)sorting was about 90%;(3)The result of RT-PCR showed that the expression level of P21 mRNA in cells infected with recombinant lentiviral was obviously de-creased,and the interferential efficiency was about 70%.Therefore,in this study,we successfully inter-fered the expression of P21 gene in the pp cells,and obtained pp cell line with decreased expression of P21 gene,which lays foundation for revealing the regulatory mechanism of P21 underlying antler regener-ation .关键词
鹿茸再生/P21 基因/RNAi/角柄骨膜/慢病毒表达载体Key words
antler regeneration/P21 gene/RNAi/pedicle periosteum/lentiviral expression vector分类
生物科学引用本文复制引用
郭倩倩,王大涛,褚文辉,鲁晓萍,秦欣,赵海平,李春义..利用慢病毒表达载体干扰梅花鹿角柄骨膜细胞P21基因[J].吉林农业大学学报,2014,(1):116-121,6.基金项目
“973”前期研究专项（2011 CB111515），吉林省自然科学基金项目 （）
