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研究川贝母表达基因的密码子的使用模式,探讨其密码子使用偏好性特点,为运用基因工程技术实现贝母碱的异源生物合成提供理论依据。以川贝母转录组9 843条全长转录本序列为数据来源,使用Codon W软件对川贝母表达基因的密码子GC含量特点、有效密码子数和同义密码子相对使用频率各项参数进行计算、统计,分析发现,川贝母表达基因密码子偏好程度整体水平不高。该研究确定了15个密码子为川贝母最优密码子UUG,CUU,AUU,GUU,UCA,CCU,CCA,ACU,ACA,GCA,UAU,CAU,AAU,AGA和GGA,最优密码子偏好A/T结尾。计算结果显示26个川贝母生物碱合成相关基因中大肠杆菌和酿酒酵母稀有密码子所占比例较低。根据稀有密码子比例,逐个分析26个关键酶基因在大肠杆菌和酿酒酵母体系中表达时可采用的密码子优化方案。该研究为通过合成生物学手段异源合成川贝母生物碱奠定了前期研究基础。
To study the codon usage patterns of Fritillaria cirrhosae gene expression and to explore the characteristics of its codon usage preference and to provide a theoretical basis for the heterologous biosynthesis of Fritillaria gmelini using genetic engineering techniques. In this study, 9 843 full-length transcripts of Fritillaria cirrhosae transcriptome were used as data sources. Codon W software was used to analyze the codon GC content, effective codon usage and synonymous codon usage frequency Calculated, statistics, analysis found that Fritillaria gene expression codon preference overall level is not high. The study identified 15 codons as the optimal codon UUG, CUU, AUU, GUU, UCA, CCU, CCA, ACU, ACA, GCA, UAU, CAU, AAU, AGA and GGA, Preferences A / T end. The results showed that the proportion of rare codons of Escherichia coli and Saccharomyces cerevisiae in 26 alkaloids of Fritillaria cirrhosa was low. According to the rare codon ratio, we can analyze the codon optimization scheme of 26 key enzyme genes expressed in E. coli and Saccharomyces cerevisiae system one by one. This study laid the foundation of previous research for the heterologous synthesis of Fritillaria cirrhosa alkaloids by means of synthetic biology.