摘要
目的:克隆夏枯草1-脱氧木酮糖-5-磷酸还原异构酶(DXR)基因并分析该基因的表达特性。方法:采用逆转录PCR技术克隆夏枯草DXR基因全长,并对其进行生物信息学分析,采用实时荧光定量PCR法分析PvDXR在不同组织及不同外源性物质诱导下的表达特性。结果:PvDXR基因编码全长为1422 bp,编码473个氨基酸组成的蛋白质序列,理论分子量为51351.49 D,为不稳定蛋白并具疏水性,含有2个跨膜螺旋区。PvDXR基因在叶中表达最高,茎中次之,果穗中表达量最少。7种外源性物质处理24 h后该基因在果穗中的表达量均有所降低,且ETH、GA3及CaCl2处理后基因的表达量降低最多。结论:获得PvDXR基因的全长cDNA,并揭示了其在不同组织及外源性物质诱导下的表达差异,为后续研究PvDXR基因在夏枯草三萜类成分合成途径中的功能奠定基础。
Objective:To clone 1-deoxyxylulose-5-phosphate reductoisomerase(DXR)gene form Prunella vulgaris and analyze its specific expression.Methods:The full-length DXR gene of Prunella vulgaris was amplified by reverse transcription PCR and bioinformatics analysis was performed.Real-time fluorescence quantitative PCR was used to analyze the expression characteristics of PvDXR induced by different tissues and different exogenous substances.Results:The PvDXR gene was encoded with a total length of 1422 bp and 473 amino acid protein sequences,its theoretical molecular weight was 51351.49 D.It was an unstable protein with hydrophobicity,contained 2 transmembrane helical regions.The expression of PvDXR gene was highest in leaf,followed by in stem,lowest in ear.When treated with seven exogenous substances for 24 h,the gene expression in the ear decreased and the ETH,GA3 and CaCl2 significantly decreased gene expression.Conclusion:The full-length cDNA of PvDXR gene is obtained,and its expression differences induced by different tissues and exogenous substances are revealed,laying a foundation for the subsequent study on the function of PvDXR gene in the synthesis pathway of triterpenoids in Prunella vulgaris.
作者
李璐
董诚明
张梦佳
朱畇昊
LI Lu;DONG Cheng-ming;ZHANG Meng-jia;ZHU Yun-hao(College of Pharmacy,Henan University of Traditional Chinese Medicine,Zhengzhou 450046,China;Collaborative Innovation Center for Respiratory Disease Diagnosis and Treatment&Chinese Medicine Development of Henan Province,Zhengzhou 450046,China)
出处
《中药材》
CAS
北大核心
2019年第6期1249-1254,共6页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金(81603232)
国家重点研发计划(2017YFC1702800)
河南省重大科技专项(171100310500)
河南中医学院博士科研基金(BSJJ2015-13)。
