Cloning and prokaryotic expression of the DIMBOA-Glc-4-OH methylase gene bx12 in maize
In order to elucidate the role of DIMBOA-Glc-4-OH methylase in the resistance of maize to aphid and mite,the gene bx12 encoding DIMBOA-Glc-4-OH methylase was cloned from four maize inbred lines with divergent resistance to aphids and tetranychids,and was prokaryotic expressed.The re-sults showed that,except for inbred line Mo17-476M,the genomes of the other three inbred lines B73,ZaC546 and Mo17-476Wwere isolated to containthe target DNA fragment of the open reading frame of the bx12 gene composed of 744 bases,and they all encoded the same protein BX12P composed of 247 amino acids.The protein was 100％consistent with the amino acid sequence of the protein encoded by the reference gene.Bioinformatics analysis revealed that BX12P is a multifunctional enzyme,belonging to the S-adenosyl-L-methionine(SAM)-dependent o-methyltransferase domain superfamily,with protein dimerization,plant methyltransferase and O-methyltransferase functions.The enzyme has the function of converting DIMBOA-Glc to HDMBOA-Glcspecifically.The target protein was successfully expressed in Escherichia coli BL21(DE3)cells using the prokaryotic expression vector constructed from cloned bx12 geneafter sequencing and structural identification.
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