Cloning of Hordein gene in barley and construction of RNAi plant expression vector.

In order to reduce the protein content and improve the quality of beer barley,this paper takes Gansu Beer Barley No.4,as the material for study,which has a wide promotion area in Gansu Province.Hordein gene was cloned in the conserved sequence,with RNAi expression vector constructed with further improvements on the theoretical basis of barley quality.A pair of primers was designed according to the released DNA sequence of Hordein gene.B2PF5'-CAACCATTTCCACAGCAACCACCAT-3',B2PR5' GAAAGATAGAGTAGACGATTGCACG-3',with PCR employed to clone a 349bp fragment(GP4) from Gansu Beer Barley No.4.Based on the structure of vector pHANNIBAL and pART27,GP4 was inserted forward and reversely according to the enzyme position.The sequence analysis shows that the cloned DNA shares 92% of identity with the data.An RNAi expression vector of the Hordein gene,named pART27-pHAN-GP4 was constructed with the RNAi expression vector pART27-pHAN-GP4 containing the object gene.