Study on Construction of shRNA Interference Plasmid by Two-step SOE PCR

摘要

A convenient RNA interference plasmid construction method was established. The shRNA interference plasmid was successfully constructed by two-step splicing overlap extention PCR (SOE-PCR). The specific interference sequence of cysteine sulfinate decarboxylase (CSD) was inserted into pEGFP-Hl-shRNA by two-step SOE-PCR. The bacterial PCR, enzyme digestion and sequence analysis results showed that the pEGFP-Hl-CSD shRNA interference plasmid were consistent with those expected. The pEGFP-Hl-CSD shRNA can be used to transfect cell and RNAi. By this method RNA interference plasmid can be rapidly and efficiently constructed which is a powerful tool for researching gene function.