| 王思懿，胡颖，刘小玉，荆欣，冯佩林，朱乐冉，姚雨桐，陈大福，郭睿，徐国钧,2023,东方蜜蜂微孢子虫LCFAL2基因的分子特性及系统进化分析[J].环境昆虫学报,45(5):1284-1290 |
| 东方蜜蜂微孢子虫LCFAL2基因的分子特性及系统进化分析 |
| Investigation of molecular characteristics and phyletic eolution of Nosema ceranae LCFAL 2 gene |
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| DOI： |
| 中文关键词: 东方蜜蜂微孢子虫 长链脂肪酸辅酶A连接酶2 分子特性 保守性 系统进化 |
| 英文关键词:Nosema ceranae long chain fatty acid COA ligase (LCFAL2) molecular characteristics conservative phyletic evolution |
| 基金项目:国家自然科学基金面上项目(32172792)；国家现代农业产业技术体系专项资金项目(CARS-44-KXJ7)；福建省自然科学基金面上项目(2022J01131334)；宁夏回族自治区重点研发项目(2022BBF02037)；福建农林大学硕士生导师团队项目(郭睿)；福建省大学生创新创业训练计划项目(202210389128, 202210389136) |
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| 中文摘要: |
| 本研究旨在解析东方蜜蜂微孢子虫Nosema ceranae的长链脂肪酸辅酶A连接酶(long chain fatty acid COA ligase)基因LCFAL2的分子特性，鉴定东方蜜蜂微孢子虫和其他物种的LCFAL2蛋白的保守基序和结构域并进行系统进化分析，以期丰富东方蜜蜂微孢子虫LCFAL2的信息，为深入开展相关功能研究提供基础。通过Expasy网站上的相关软件预测和分析LCFAL2的理化性质、跨膜螺旋域、信号肽、磷酸化位点、二级结构和三级结构。使用MEME软件预测东方蜜蜂微孢子虫和其他物种LCFAL2蛋白的保守基序。采用TBtools软件预测东方蜜蜂微孢子虫和其他物种LCFAL 2的结构域。通过Mega 11.0软件构建东方蜜蜂微孢子虫和其他物种的LCFAL 2的系统进化树。结果表明，东方蜜蜂微孢子虫LCFAL 2含有1 836个核苷酸，可编码611个氨基酸，分子式为C3145H4985N813O904S23，分子量约为69.39 kDa，脂溶系数为96.82，等电点为8.79，平均亲水系数为-0.161；不含跨膜螺旋域和典型信号肽；包含27个丝氨酸酸化位点，14个酪氨酸酸化位点及23个苏氨酸磷酸化位点；包含240个α-螺旋，124个延长链，48个β-转角及199个无规则卷曲；可同时定位于细胞核、细胞质和线粒体；另外，LCFAL2及其模板5mst.1.A之间的同源性为21.44%。东方蜜蜂微孢子虫与蜜蜂微孢子虫Nosema apis等9个物种的LCFAL 2中均含有6个相同的保守基序。东方蜜蜂微孢子虫、管孢属微孢子虫Tubulinosema ratisbonensis、蜜蜂微孢子虫、巨蟹肠孢菌Enterospora canceri、烟粉虱Spraguea lophii、毕氏肠细胞虫Enterocytozoon bieneusi、东方赤孢子虫Hamiltosporidium tvaerminnensis和褶皱臂尾轮虫Brachionus plicatilis的LCFAL 2均包含AFD_class_I superfamily结构域。东方蜜蜂微孢子虫LCFAL2(XP_024331795.1)与蜜蜂微孢子虫LCFAL2(EQB61677.1)聚为一支，进化距离最近。研究结果明确了东方蜜蜂微孢子虫LCFAL2的分子特性，并揭示LCFAL2可能是亲水性蛋白、胞内蛋白和非跨膜蛋白，东方蜜蜂微孢子虫与蜜蜂微孢子虫的LCFAL2之间的亲缘关系最近，LCFAL2在东方蜜蜂微孢子虫和其他微孢子虫中具有较高的保守性。 |
| 英文摘要: |
| This study aims to decipher the molecular characteristics of long chain fatty acid COA ligase 2 gene LCFAL2 in Nosema ceranae, identify conserved motifs and structural domains and conduct phylogenetic analysis of LCFAL 2 proteins in N. ceranae and other species, thus enriching information about N. ceranae LCFAL2 gene and providing a foundation for related functional study. Physical and chemical property, transmembrane domain, signal peptide, phosphorylation site, secondary and tertiary structures were predicted and analyzed using relevant software on Expasy website. MEME software was emplyed to identify conserved motifs within LCFAL2 in N. ceranae and other species. Conserved domains within LCFAL2 were predicted with TBtools software. Mega 11.0 software was utilized to construct the phylogenetic tree with neighbor-joining method based on LCFAL2 proteins in N. ceranae and other species. The results demonstrated that N. ceranae LCFAL2 contained 1 836 nucleotides, which can encode 611 amino acids, the molecular formula was C3145H4985N813O904S23, the molecular weight was about 69.39 kDa, the lipid solubility coefficient was 96.82, the isoelectric point was 8.79, and the average hydrophilic coefficient was -0.161; LCFAL2 contained no transmembrane domain and signal peptide; LCFAL2 included 27 serine phosphorylation sites, 14 tyrosine phosphorylation sites and 23 threonine phosphorylation sites; LCFAL2 included 240 α-helix, 124 extension, 48 β-turn and 199 random coil; the homology between LCFAL2 and its target 5mst.1.A was 21.44%; additionally, LCFAL2 were simultaneously localized in nucleus, cytoplasm and mitochondria. Six same conserved motifs were included in LCFAL2 proteins in nine species such as N. ceranae and N. apis. Also, AFD_class_I superfamily structural domain were identified in LCFAL2 in N. ceranae, Tubulinosema ratisbonensis, N. apis, Encephalitozoon cuniculi, Spraguea lophii, Enterocytozoon bieneusi, Hamiltosporidium tvaerminnensis and Brachionus plicatilis. LCFAL2 (XP_024331795.1) in N. ceranae and LCFAL2 (EQB61677.1) were clustered into one clade, and their evolutionary distance was the closest. These findings clarify the molecular characteristics of N. ceranae LCFAL2, and uncover that LCFAL2 may be hydrophilic protein, intracellular protein and non-transmembrane protein, the genetic relationship between N. ceranae LCFAL2 and N. apis LCFAL2 is the closet, and LCFAL2 is highly conserved in N. ceranae and other microsporidia. |
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