Cloning and sequencing of OsMAPK4 gene and construction of its plant expression vector

A 1 500 bp DNA fragment amplified by RT-PCR use OsMAPK4 gene special primer from liaoyan241 leaf treated by low temperature was cloned into pUC18. The result of sequencing showed that the sequence of this fragment was 99.4% homologous with that of OsMAPK4 gene in GenBank and amino acid homologous is 99.1%. Further more OsMAPK4 gene was cloned into plant express vector cassette pBCE12 and pBC29A. Thus two plant express vectors, pBME12 and pBM29A were constructed, in which OsMAPK4 gene was controlled 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 OsMAPK4 gene into plant by Agrobacterium-mediated method and its function analysis.