[关键词]
[摘要]
目的 从甘草Glycyrrhiza uralensis中克隆1个新的NADPH细胞色素P450还原酶基因(GuCPR,登录号:MH401048)并进行生物信息学分析。方法 提取甘草根部总RNA后逆转录为cDNA,通过转录组数据库筛选得到GuCPR基因全长,利用NCBI ORF finder得到其开放阅读框并翻译得氨基酸序列,设计引物进行PCR扩增,构建克隆重组质粒。利用生物信息分析预测蛋白性质、结构等及模拟蛋白三级结构,并构建同源系统进化树。结果 克隆得到GuCPR基因cDNA全长2 118 bp,编码705个氨基酸残基,相对分子质量78 450,等电点(pI)5.19。GuCPR蛋白为非分泌蛋白,不具有信号识别功能。跨膜预测结果显示蛋白的第44~64位氨基酸为跨膜区,同时,亚细胞定位预测结果显示蛋白位于内质网上。结论 克隆得到一个新的GuCPR,并进行了生物信息学分析,为进一步研究提供了理论基础。
[Key word]
[Abstract]
Objective To clone a new NADPH cytochrome P450 reductase gene (GuCPR) (Login number:MH401048) from Glycyrrhiza uralensis and do some bioinformatics analysis. Methods Total RNA was extracted from the roots of G. uralensis and then transcription reversed into cDNA. The GuCPR gene was obtained by screening the transcriptome database. The NCBI ORF finder was used to obtain its open reading frame and translated amino acid sequence, design primers for PCR amplification, construction of recombinant cloned plasmid. Bioinformatics analysis was used to predict the protein properties, structure and model the tertiary structure of the protein, and homologous phylogenetic tree construction was performed. Results cDNA of GuCPR gene was cloned to a total length of 2 118 bp and encoded 705 amino acid residues with a relative molecular mass of 78 450. Electricity (pI) 5.19. GuCPR protein was a non-secretory protein and did not have a signal recognition function. The results of transmembrane prediction showed that the amino acids 44-64 of the protein were transmembrane regions. Meanwhile, the subcellular localization prediction results showed that the protein was located on the endoplasmic reticulum. Conclusion A new GuCPR was cloned and its bioinformatics analysis laid the foundation for further research.
[中图分类号]
R282.12
[基金项目]
国家自然科学基金资助项目(81773838)