The cloning system based on RNAi GG vector construct using type IIS restriction enzyme BsaI is deviced.

Vector containing a Pdk intron region; form a hairpin loop structure, two copies of the ccdB gene flanked by BsaI restriction sites. For SaLac1 gene insertion to the RNAi-GG vector, the ccdB genes are removed with BsaI; identifying sequences upstream and downstream of the pdk intron. Subsequently; adaptor sequences specific to the RNAi-GG vector are then added to Salac gene, enabling its insertion in both sense and antisense directions on either side of the Pdk intron region. The resulting hairpin loop incorporates Salac; triggers the SaLac silencing through RNA interference.
(DNA)