Cloning and sequencing of OsMAPK7 gene and construction of its plant expression vector

A 1 700 bp DNA fragment amplified by RT-PCR use OsCDPK7 gene special primer from liaoyan241 leaf treated by 200 mmol·L-1 NaCl was cloned into pUC18.The result of sequencing showed that this fragment 78 nucleotides from 157-234 in CDS compare with that of OsCDPK7 gene in GenBank and lack of 26 amino acids from 53-78 in protein sequence.Further more,OsCDPK7 gene was cloned into plant express vector cassette pBE12 and pB29A.Thus two plant express vectors,pBC7E12 and pBC729A were constructed,in which OsCDPK7 gene was regulated by E12 promoter and rd29A promoter respectively.Then the recombination plasmids were introduced into Agrobacterium LBA4404 by freezing-melting transformation method.This work provides a foundation for transferring OsCDPK7 gene into plant by Agrobacterium-mediated method and its function analysis.