蒙古黄芪异黄酮合成酶基因密码子偏好性分析
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摘要
异黄酮是只局限于豆科蝶形花亚科等极少数植物中的一种植物性雌激素,异黄酮合成酶(IFS)是苯丙氨酸分支代谢途径——异黄酮代谢途径的关键酶。为了解蒙古黄芪IFS基因密码子的使用特性,采用CodonW,SPSS软件及EMBOSS在线程序分析蒙古黄芪IFS基因密码子的偏好性,并分别与14个物种的IFS以及模式生物基因组进行比较。结果显示,蒙古黄芪IFS基因的密码子选择偏性较弱且较偏好以A/T结尾,而物种间IFS密码子选择偏性则存在一定差异;进化树分析表明,基于IFS编码序列聚类结果比密码子使用偏性分类结果能更准确地反映物种间的亲缘关系;密码子使用频率比较结果发现,酵母真核表达系统更适用于蒙古黄芪IFS基因异源表达系统建立,而蒙古黄芪IFS基因与模式植物基因组之间密码子使用偏性差异较小,尤其拟南芥可能为该基因转基因研究最为理想的受体。
Isoflavone is a kind of plant estrogens which are restricted to only a few plants in the family Leguminosae. Isoflavone synthetase is the key enzyme of isoflavone synthesis pathway-Phyenlalanine metabolism branch pathway. To understand the codon bias of the IFS gene, one IFS gene of Astragalus. membranaceus var. mongholicus was analyzed by CodonW, SPSS software and EMBOSS online program, and compared with the 14 species IFS and the genomes of model organisms, respectively. The results showed that the selection bias of the IFS gene of Astragalus. membranaceus var. mongholicus was weaker, they were bias toward the synonymous codons with A and T at third codon position. Phylogenetic tree analysis showed that the clustering results based on IFS coding sequence could reflect the phylogenetic relationship among species more accurately than the result analyzed by RSCU(Relative Synonymous Codon Usage). It was found that the yeast eukaryotic expression system was more suitable for the establishment of the heterologous expression system of the IFS gene of Astragalus. membranaceus var. mongholicus, and the difference of codon bias between the IFS gene of Astragalus.membranaceus var. mongholicus and the genomes of the pattern plant were not significant, especially the Arabidopsis thaliana may be the most ideal receptor for the gene transfer.
Isoflavone is a kind of plant estrogens which are restricted to only a few plants in the family Leguminosae. Isoflavone synthetase is the key enzyme of isoflavone synthesis pathway-Phyenlalanine metabolism branch pathway. To understand the codon bias of the IFS gene, one IFS gene of Astragalus. membranaceus var. mongholicus was analyzed by CodonW, SPSS software and EMBOSS online program, and compared with the 14 species IFS and the genomes of model organisms, respectively. The results showed that the selection bias of the IFS gene of Astragalus. membranaceus var. mongholicus was weaker, they were bias toward the synonymous codons with A and T at third codon position. Phylogenetic tree analysis showed that the clustering results based on IFS coding sequence could reflect the phylogenetic relationship among species more accurately than the result analyzed by RSCU(Relative Synonymous Codon Usage). It was found that the yeast eukaryotic expression system was more suitable for the establishment of the heterologous expression system of the IFS gene of Astragalus. membranaceus var. mongholicus, and the difference of codon bias between the IFS gene of Astragalus.membranaceus var. mongholicus and the genomes of the pattern plant were not significant, especially the Arabidopsis thaliana may be the most ideal receptor for the gene transfer.
引文
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[18]晁岳恩,吴政卿,杨会民,等.11种植物psbA基因的密码子偏好性及聚类分析[J].核农学报,2011,25(5):927-932.
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[21]罗火林,张亚楠,罗丽萍,等.兰科植物FT基因的密码子偏性分析[J].分子植物育种,2016,14(1):51-58.
[22]吴芳明.拟南芥剪切因子SKIP参与生物钟调控的研究[D].北京:北京师范大学,2011.
[2]郝占西.大肠杆菌亮氨酸t RNA毫秒级构象变化的NMR研究[D].上海:中国科学院生物化学与细胞生物学研究所,中国科学院上海生物化学和细胞生物学研究所,中国科学院上海生命科学研究院生物化学与细胞生物学研究所,中国科学院上海生命科学研究院,2010.
[3]谢平.遗传密码子的起源——从能量转化到信息化[J].生物多样性,2017,25(1):94-106.
[4]高爱琴,李宁,李金泉,等.山羊FGF5基因c DNA分子克隆及序列分析[J].内蒙古大学学报(自然科学版),2006,37(2):180-184.
[5]范新阳,邱立华,罗国祥,等.河流型与沼泽型水牛CSN3基因密码子使用特征分析[J].云南农业大学学报(自然科学),2017,32(2):238-245.
[6]陈义挺,赖瑞联,冯新,等.猕猴桃查尔酮合成酶基因CHS密码子偏好性分析[J].福建农业学报,2017(11):1213-1217.
[7]续晨,贲爱玲,蔡晓宁.蝴蝶兰叶绿体基因组密码子使用的相关分析[J].分子植物育种,2010,8(5):945-950.
[8]韩淑晓,孙福来,刘全兰,等.植物抗病基因密码子使用特性分析[J].山东农业大学学报(自然科学版),2013,44(4):491-498.
[9]胡艳改,杜宇忠,郝磊,等.中药材蒙古黄芪与膜荚黄芪优质种源的研究[J].山西农业科学,2017,45(10):1662-1666,1690.
[10]任杰,梁建萍,周然,等.不同光质对黄芪生长及药用成分积累的影响[J].山西农业科学,2014,42(10):1078-1081.
[11]王栋,李安平,王玉龙,等.蒙古黄芪种子质量检验方法研究[J].山西农业科学,2015,43(4):460-464.
[12]宋奇.中药黄芪在驻颜抗衰老方面的研究进展[J].中国新技术新产品,2011(8):10-12.
[13]罗建平,王军辉,范远景.植物异黄酮合酶研究进展[J].中国生物工程杂志,2005,25(7):17-20.
[14]张来,杨碧昌,刘宁.植物黄酮类化合物在医药上的应用[J].安顺学院学报,2010,12(2):84-88.
[15]JIANG N,JEON E H,PAK J H,et al.Increase of isoflavones in soybean callus by Agrobacterium-mediated transformation[J].Plant Biotechnology Reports,2010,4(4):253-260.
[16]FUGLSANG A.The effective number of codons for individual amino acids:some codons are more optimal than others[J].Gene,2003(3):185-190.
[17]刘汉梅,赵耀,顾勇,等.几种植物waxy基因的密码子用法特性分析[J].核农学报,2010,24(3):476-481.
[18]晁岳恩,吴政卿,杨会民,等.11种植物psbA基因的密码子偏好性及聚类分析[J].核农学报,2011,25(5):927-932.
[19]MURRAY E E,LOTZER J,EBERLE M.Codon usage in plant genes[J].Nucleic Acids Research,1989,17(2):477.
[20]CHRISTIANSON D W.Arginase:Structure,mechanism,and physiological role in male and female sexual arousal[J].Cheminform,2005,36(24):191-201.
[21]罗火林,张亚楠,罗丽萍,等.兰科植物FT基因的密码子偏性分析[J].分子植物育种,2016,14(1):51-58.
[22]吴芳明.拟南芥剪切因子SKIP参与生物钟调控的研究[D].北京:北京师范大学,2011.