Molecular cloning of gene fragment encoding 4-coumarate: Coenzyme A ligase of Sengon (Paraserianthes falcataria)

Abstract

4-coumarate:Coenzyme A ligase (4CL) plays an important role in lignin biosynthetic pathway that catalyzed the activation of coumaric acid, caffeic acid or ferulic acid to be a syringil monomer. Lignin biosynthesis control through 4CL down regulating would support lower lignin wood production. The objective of this study was to clone conserved region cDNA of gene encoding 4CL. Gene fragment isolation was conducted by means of reverse transcriptase polymerase chain reaction (RT-PCR) using degenerate heterologous primer. The RT-PCR products were purified, sequenced and analyzed to select the highly homologous fragment to 4CL. BLASTanalysis result showed that deduction of amino acid sequences from one of two RT-PCR product nucleotide was highly homologous with the 4CL conserved region from Rubbus ideaus, Oryza sativa, Populus tomentosa, Populus balsamifera, Betulla platyphilla, Nicotiana tabacum, and Arabidopsis thaliana with identity ranging from 78-90%. Key words: 4-coumarate: Coenzyme A ligase, lignin, sengon