中国农业大学学报2024，Vol.29Issue(4)：195-204,10.DOI:10.11841/j.issn.1007-4333.2024.04.17
枸杞查耳酮异构酶LcCHI1基因的克隆与表达分析
Cloning and expression analysis of Lycium chinense chalcone isomerase LcCHI1 gene
摘要
Abstract
The expression patterns and functions of chalcone isomerase CHI1 gene from different tissues in Lycium chinense were investigated in this study.LcCHI1 gene(Genbank No.OR026273)was obtained using homologous cloning method.Its physicochemical properties,structure,and evolutionary relationships were analyzed using bioinformatics methods.The tissue expression specificity was analyzed using fluorescence quantitative PCR(qRT-PCR)technology and prokaryotic expression analysis was performed.The results showed that:1)The LcCHI1 gene had a total length of 695 bp,an open reading frame of 633 bp,encoding 210 amino acids and containing a conserved chalcone_3 family domains.2)Multiple sequence alignment showed that the LcCHI1 protein conformed to the structural characteristics of chalcone protein and had high similarity with other chalcone proteins.Cluster analysis showed that the LcCHI1 protein and the CHI protein of the Solanaceae family aggregated into a single branch.3)LcCHI1 was highly expressed in the flowers,tender leaves,and tender branches of L.chinense,but low in the roots.With the fruit development of L.chinense,the expression of LcCHI1 showed a trend of first increasing and then decreasing,with the highest expression in color changing fruits.4)The LcCHI1 recombinant protein in the prokaryotic expression vector was constructed,and the expressed protein appeared at 63 ku through SDS-PAGE electrophoresis analysis.In summary,the LcCHI1 gene was highly expressed in flowers and color changing fruits from L.chinense,and can undergo prokaryotic protein expression.The results layed the foundation for the functional verification of the CHI gene in the flavonoid synthesis pathway of L.chinense.关键词
枸杞/查耳酮异构酶/基因克隆/基因表达/原核表达Key words
Lycium chinense/chalcone isomerase/gene cloning/gene expression/prokaryotic expression分类
生物科学引用本文复制引用
乔枫..枸杞查耳酮异构酶LcCHI1基因的克隆与表达分析[J].中国农业大学学报,2024,29(4):195-204,10.基金项目
青海省共和县黄河龙羊峡库区林草生态修复综合治理中央财政支持国土绿化示范试点项目(科技支撑)(E6301000076033516) （科技支撑）
中藏药材绿色高值化与标准化生产技术集成与示范推广项目(2023-NK-P38) （2023-NK-P38）
