Construction and identification of specific shRNA interference plasmid vector targeted to DAL-1 gene mRNA
AIM: To build the experimental NSCLC cellular model in which expression of DAL-1 is steadily inhibited. METHODS: Specific shRNA interference plasmid vectors targeted to DAL-1 mRNA was constructed. The positive clones were screened by double-enzyme digestion and DNA sequencing. The expression at levels of DAL-1 mRNA and protein was detected with Q-PCR and Western blot to screen the clones in which DAL-1 was steady inhibited. RESULTS: The sequences of the constructs were confirmed by double-enzyme digestion and DNA sequencing. The expressions of DAL-1 mRNA and DAL-1 protein of T4 group decreased significantly and the inhibition rate was(87.4 ±2.0)% and (82. 7 ±2. 1)% respectively. CONCLUSION: Specific shRNA interference plasmid vector targeted to DAL-1 gene mRNA is successfully designed and constructed. NCI-H460/T cell line with stable inhibition of DAL-1 expression is established successfully , which provides experimental models for further study of the roles of DAL-A in NSCLC cells.
NETL
NSTL
万方数据
维普
