Identification and Properties of Ketoacyl-coenzyme A Synthetase Gene in Primulina spinulosa
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摘要:
目的 为进一步探讨刺齿报春苣苔酮脂酰-辅酶A合成酶基因的分子结构和功能，并为刺齿报春苣苔适应极端干旱环境提供分子依据。 方法 总RNA的提取采用改良Trizol法，通过转录组测序获得与刺齿报春苣苔（Primulina spinulosa）抗旱及应对生物胁迫反应有关的基因：酮脂酰-辅酶A合成酶基因，并运用DNAman、Finder、TMPRED、SWISS-MODEL、NetPhos2.0、SignaIP4.1等软件对序列的开放阅读框（ORF）、编码氨基酸、编码蛋白质等理化特性进行分析，并构建系统发育树。 结果 序列分析结果表明，该基因全长1 935 bp（GenBank登录号为MH543314），开放阅读框（ORF）为1 599 bp，共编码532个氨基酸，编码蛋白质的分子量为59.82 kDa，理论等电点（pI）为9.16，为亲水性蛋白，含7个跨膜区，30个可能的磷酸化位点，1个与查尔酮合成酶家族蛋白相同的保守结构域；它与同科旋蒴苣苔（Dorcoceras hygrometricum，KZV37788）的同源性高达92%，两者在系统进化树上聚为一支。 结论 酮脂酰-辅酶A合成酶基因与刺齿报春苣苔叶表面的蜡质合成有关，对植物的抗旱及应对生物胁迫发挥重要调控作用。 -
关键词:
- 刺齿报春苣苔 /
- 酮脂酰-辅酶A合成酶基因 /
- 基因序列分析
Abstract:Objective The ketoacyl-coenzyme A synthase (KCAS) gene relating to responses of Primulina spinulosa to drought and other stresses was studied based on the transcriptome sequencing. Method Total RNA of the gene was extracted by the improved Trizol method for sequencing with the established database. Physicochemical properties of the open reading frame (ORF), coding amino acids, and proteins of the gene were analyzed by DNAman, Finder, TMPRED, SWISS-MODEL, NetPhos 2.0, and SignaIP4.1. The phylogenetic tree was constructed accordingly. Results The full length of the KCAS gene was determined to be 1 935 bp (GenBank login number MH543314), ORF 1 599 bp, encoding 532 amino acids, molecular weight 59.82 kDa, and theoretical pI 9.16. It was a hydrophilic protein with 7 transmembrane regions, 30 possible phosphorylation sites, and one conservative domain similar to that of the chalcone synthase family. The homology between it and Dorcoceras hygrometricum (KZV788) was as high as 92%, and they were in a same phylogenetic tree. Conclusion The KCAS gene was postulated to relate to the leaf wax synthesis of P. spinosa and played an important role in drought resistance and biological response to stresses of the plant. -
图 1 酮脂酰-辅酶A合成酶基因序列和推测的氨基酸序列
注：开放阅读框从氨基酸(M)开始至终止密码(*)止。
Figure 1. Sequence of KCAS gene and deduced amino acid sequence
Note: Open reading frame starts from amino acid (M) to termination code (*)
图 4 刺齿报春苣苔酮脂酰-辅酶A合成酶基因编码蛋白的跨膜区预测
Figure 4. Predicted transmembrane region of protein encoding KCAS gene in P. spinulosa
图 7 SWISS-Model模拟的刺齿报春苣苔酮脂酰-辅酶A合成酶蛋白的三级结构
Figure 7. Tertiary structure of KCAS protein of P. spinulosa simulated by SWISS-Model
图 8 基于氨基酸序列的酮脂酰-辅酶A合成酶系统发育树
注：Spi|c32695_g1(刺齿报春苣苔，MH543314)，Dorcoceras hygrometricum(旋蒴苣苔，KZV37788)，Sesamum indicum(芝麻，XP_011081956)，Erythranthe guttata(水曲柳，XP_012855831)，Olea europaea var. Sylvestris(油橄榄，XP_022868534)，Theobroma cacao(可可，EOY30095)，Helianthus annuus(向日葵，XP_021982926)，Cynara cardunculus var. Scolymus(菜蓟，XP_024961571)，Gossypium hirsutum(棉花，NP_001313740)，Coffea canephora(中果咖啡，CDP17887)，Ipomoea nil(牵牛花，XP_019167964)，Herrania umbratica(乌骨草，XM_021420693)，Ziziphus jujuba(枣，XP_015884557)。
Figure 8. Phylogenetic tree of KCAS based on amino acid sequence
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