香椿CCR家族基因TsCCR1的克隆与生物信息学分析
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摘要
肉桂酰辅酶A还原酶在木质素生成过程中发挥重要作用,本研究从香椿幼苗中克隆得到1个肉桂酰辅酶A还原酶家族基因(CCR)。生物信息学分析表明,该基因含有924 bp的开放阅读框,共编码307个氨基酸,分子量为35.48kDa,等电点为5.34,亲水性平均系数为-0.162,为亲水性、酸性蛋白;蛋白结构域分析发现该基因编码的蛋白含有底物结合位点、酶活性位点以及NADP结合位点,属于SDR超家族成员;系统进化树分析表明,该基因编码的蛋白与胡杨CCR1的亲缘关系最近,因此将其命名为TsCCR1。其基因的克隆为通过基因工程手段改良香椿优良品种的栽培抗性提供了参考。
Cinnamoyl-CoA reductase plays a major role in lignin biosynthesis. In this study, one of cinnamoyl-CoA reductase genes(CCR) was cloned from Toona sinensis seedlings. Bioinformatics analysis showed that the gene contained an open reading frame of 924 bp which encoded a total of 307 amino acids with molecular weight of 35.48 kDa, isoelectric point of 5.34 and grand average of hydropathicity of-0.162. The protein is hydrophilic and acidic. Protein domain analysis revealed the protein contained substrate binding sites, enzyme active sites and NADP binding sites, belonged to the SDR super-family. Phylogenetic tree analysis showed the protein was closely related to Populus euphratica CCR1, so it is named TsCCR1. The TsCCR1 gene provides a reference for improving Toona sinensis varieties resistance through genetic engineering.
Cinnamoyl-CoA reductase plays a major role in lignin biosynthesis. In this study, one of cinnamoyl-CoA reductase genes(CCR) was cloned from Toona sinensis seedlings. Bioinformatics analysis showed that the gene contained an open reading frame of 924 bp which encoded a total of 307 amino acids with molecular weight of 35.48 kDa, isoelectric point of 5.34 and grand average of hydropathicity of-0.162. The protein is hydrophilic and acidic. Protein domain analysis revealed the protein contained substrate binding sites, enzyme active sites and NADP binding sites, belonged to the SDR super-family. Phylogenetic tree analysis showed the protein was closely related to Populus euphratica CCR1, so it is named TsCCR1. The TsCCR1 gene provides a reference for improving Toona sinensis varieties resistance through genetic engineering.
引文
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[10]隋娟娟,孙晶晶,杨京霞,等.香椿TsCCR基因的克隆及表达分析[J].分子植物育种,2018,16(17):5567-5575.
[11]吴蓓,李梦瑶,王广龙,等.芹菜肉桂酰辅酶A还原酶基因的克隆与表达分析[J].南京农业大学学报,2016,39(6):907-914.
[12]李波,梁颖.柴友荣.植物肉桂酰辅酶A还原酶(CCR)基因的研究进展[J].分子植物育种,2006,4(3S):55-65.
[2]唐晓珍,王明林,乔聚林,等.香椿的营养与保健作用[J].中国食物与营养,2003,(2):48-49.
[3]杨京霞,李静,吴婷,等.太和香椿水提物对秀丽隐杆线虫生物学特征的影响及清除自由基作用的研究[J].食品工业科技,2016,37(11):328-331.
[4]DOWD P F,SARATH G,MITCHELL R B,et al.Insect resistance of a full sib family of tetraploid switchgrass Panicum virgatum L.with varying lignin levels[J].Genetic Resources&Crop Evolution,2013,60(3):975-984.
[5]耿飒,徐存拴,李玉昌.木质素的生物合成及其调控研究进展[J].西北植物学报,2003,23(1):171-181.
[6]董丽丽,龚凌燕,陈磊,等.石榴肉桂酰辅酶A还原酶基因的克隆与表达分析[J].南京农业大学学报,2016,39(5):747-753.
[7]LEPLéJ C,GRIMA-PETTENATI J,VAN MON-TAGU M,et al.A cDNA encoding cinnamoyl CoA reductase from Populus trichocarpa[J].Plant Physiology,1998,117(3):1126.
[8]PIQUEMAL J,LAPIERRE C,MYTON K,et al.Down-regulation of Cinnamoyl-CoA Reductase induces significant changes of lignin profiles in transgenic tobacco plants[J].Plant Journal,1998,13(1):71-83.
[9]李高,杨杞,张烨,等.柠条锦鸡儿肉桂酰辅酶A还原酶基因克隆和分析[J].中国生物工程杂志,2014,34(1):50-56
[10]隋娟娟,孙晶晶,杨京霞,等.香椿TsCCR基因的克隆及表达分析[J].分子植物育种,2018,16(17):5567-5575.
[11]吴蓓,李梦瑶,王广龙,等.芹菜肉桂酰辅酶A还原酶基因的克隆与表达分析[J].南京农业大学学报,2016,39(6):907-914.
[12]李波,梁颖.柴友荣.植物肉桂酰辅酶A还原酶(CCR)基因的研究进展[J].分子植物育种,2006,4(3S):55-65.