Cloning and Bioinformatics Analysis of GI4CL Gene in Glehnia littoralis / 世界科学技术-中医药现代化

ABSTRACT

This study was aimed to clone and analyze the open reading frame (ORF) of 4-coumarate:coenzyme A ligase (Gl4CL) gene in Glehnia littoralis.Based on the high-throughput sequencing of G.littoralis,the full-length cDNA of Gl4CL gene was cloned by the rapid amplification of cDNA ends (RACE) method.Physical and chemical properties,secondary structure and three-dimensional structure of Gl4CL protein were predicted.Real-time PCR was used to detect the expression of Gl4CL gene in roots and leaves of G.littoralis.A total of 1951 bp full-length cDNA of Gl4CL gene was obtained,which encoded a protein of 544 amino acids with a predicted molecular weight of 59.481 kDa and the isoelectric point of 8.20.The cDNA of Gl4CL gene included 1 635 bp of ORF,153 bp of 5'untranslated regions (5'UTR) and 163 bp of 3'UTR.The result of real-time PCR showed that Gl4CL gene was both expressed in roots and leaves of G.littoralis,while the expression of gene in roots was significantly higher than that in leaves.It was concluded that the study will lay the foundation for further study of Gl4CL gene in function and gene regulation.Through in-depth study of the relationship between the expression of Gl4CL gene and lignin,as well as the plant growth phenotypes,it is expected to obtain high yield and quality lines of Glehniae Radix with strong resistance to diseases and insect pests.