Cloning of Caffeoyl-coenzyme A O-methyltransferase Gene from Citrus maxima (Burm.) Merr. and Its Plant Expressing Vector Construction

doi:10.11924/j.issn.1000-6850.2014-0400

Abstract

Abstract: To probe the effects of the caffeoyl-coenzyme A 3-O-methyltransferase on lignin biosynthesis in granulated pummelo juice sac, a cmCCoAOMT gene was cloned from pummelo juice sac and constructed into plant expressing vector, and then was transformed into Agrobacterium tumefaciens. Total RNA was obtained by Trizol method, and was reverse transcribed as cDNA. cmCCoAOMT gene was cloned by PCR with a primer pair designed from a CCoAOMT gene sequence of ORF. The open reading frame (ORF) contained 744 base pairs coding 247 amino acids. The sequence homology with Populus balsamifera, Jatropha curcas and Dimocarpus longan exceeded 90% . Prediction result indicated that cmCCoAOMT was located on ER with possibility of 0.595 and it was not a transmembrane protein. The over expression vector obtained by inserting the ORF of cmCCoAOMT cDNA into p1301 under the control of 35S promoter was transformed into Agrobacterium tumefaciens. As a conclusion, the ORF of cmCCoAOMT gene was cloned and constructed into plant expression vector p1301 in this study. Agrobacterium tumefaciens contained the vector can infect many plants and be used as the base for gene function study.

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