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

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.

Analysis on codon using features and expression prediction of 4CL gene in Dryopteris fragrans / 中草药

Objective: Expression and analysis of 4CL gene of ferns, which lays the theoretical foundation for the accumulation and regulation of secondary metabolites of ferns. Methods: Cloned 4CL gene (Df4CL) sequence from Dryopteris fragrans was analyzed by CHIPS, CUSP, and Codon W programs online, and then compared with Escherichia coli, yeast, Arabidopsis thaliana, and Nicotiana tabacum L. It is important to identify the codon usage of Df4CL gene and select appropriate expression systems. Results: Df4CL gene had not the bias toward the synonymous codons with ATGC at the third codon position, but had the preference consistency with Physcomitrella patens codon usage. The differences in codon usage frequency between Df4CL gene and A. thaliana were less than those in N. tobacum, E. coli, and yeast. Based on 4CL gene codon usage bias of cluster analysis, it was shown that D. fragrans, P. patens, and A. thaliana were classed into one group. Conclusion: The A. thaliana expression system may be more suitable for the exogenous expression of Df4CL gene of D. fragrans.