黄芩查尔酮异构酶基因的全长cDNA克隆及生物信息学分析

Cloning and Bioinformatic Analysis of Chalcone Isomerase Gene Full‑length cDNA from Scutellaria baicalensis

目的:从黄芩叶片克隆全长查尔酮异构酶基因chi并进行功能鉴定。方法:从黄芩叶片mRNA中RT-PCR获得全长chi基因并进行生物信息学分析。结果:克隆的chi基因全长648 bp,含有完整chi基因的开放阅读框,拟编码215个氨基酸,具有查尔酮异构酶催化特有的结构域PLN02559,属于查尔酮_3超家族。构建表达载体pET-32a(+)-chi,在IPTG诱导下在大肠杆菌中表达出一个30.0 kDa的融合蛋白。结论:分析表明,黄芩叶片chi与从黄芩组培毛状根、愈伤组织的chi基因序列几乎完全相同,预示黄芩叶片可替代野生黄芩根用药,为野生黄芩资源的可持续利用提供理论依据。

Objective:To clone a full‑length chi from scutellaria baicalensis leaves and identify its func‑tion.Methods:One full-length chi was amplified from S.baicalensis leaves mRNA by RT-PCR and ana‑lyzed by bioinformatics.Results:The cloned chi was 648 bp,contained a complete open reading frame of chi which encoding 215 amino acids,contained a unique catalyzed domain PLN02559 of chalcone isom‑erase,and belonged to chalcone_3 superfamily.The expression vector pET-32a(+)-chi was constructed and a 30.0 kDa fusion protein was expressed in Escherichia coli which induced by IPTG.Conclusion:The chi sequence in S.baicalensis leaves had the same as these in hairy roots and callus,and S.baicalensis leaves could be as substitutes of S.baicalensis root used in Chinese medicine.It can provide theoretical basis for the sustainable utilization of wild S.baicalensis resources.