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| 东方蜜蜂微孢子虫蓖麻毒素B凝集素基因的分子克隆及生物信息学分析 |
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范元婵1,王杰1,孙明会1,吴鹰1,余岢骏1,王心蕊1,叶亚萍1,钱加珺1,张凯遥1,王思懿1,陈大福1,2,郭睿1,2*
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| (1.福建农林大学 动物科学学院(蜂学学院), 福州 350002;2.福建农林大学 蜂疗研究所, 福州 350002) |
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| 摘要: |
| 为获得东方蜜蜂微孢子虫(Nosema ceranae)的NCRBL-081基因序列，并对NCRBL-081编码蛋白进行预测和分析。本研究利用RT-PCR克隆NCRBL-081基因，利用相关生物信息学工具对NCRBL-081编码蛋白进行预测。结果表明:NCRBL-081共含有630个核苷酸，可编码含209个氨基酸的亲水性蛋白；NCRBL-081编码蛋白含有1个信号肽，6个明显的丝氨酸，5个酪氨酸磷酸化位点及1个跨膜结构域；该蛋白含有20.10%的α螺旋，8.61% 的β折叠，25.84%的延伸链及45.45%的无规卷曲，且具有Ⅱ型核糖体失活蛋白家族的典型特征；同一个微孢子虫物种的不同RBL蛋白之间进化距离较近，而不同微孢子虫物种的RBL蛋白之间进化距离较远；东方蜜蜂微孢子虫的NCER_100581与AAJ76_100008081之间亲缘关系较近。研究结果解析了东方蜜蜂微孢子虫NCRBL-081的序列，并揭示了东方蜜蜂微孢子虫NCRBL-081编码蛋白的分子特征，可为后续功能探究提供依据。 |
| 关键词: 东方蜜蜂微孢子虫 蓖麻毒素B凝集素 分子克隆 生物信息学 系统进化树 |
| DOI：10.11841/j.issn.1007-4333.2022.03.15 |
| 投稿时间：2021-04-28 |
| 基金项目:国家现代农业产业技术体系建设专项资金(CARS-44-KXJ7)；福建农林大学杰出青年科研人才计划项目(xjq201814)；江西省蜜蜂生物学与饲养重点实验室开放基金项目(JXKLHBB-2020-04) |
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| Molecular cloning and bioinformatic analysis of Nosema ceranae ricin B-lectin gene |
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FAN Yuanchan1,WANG Jie1,SUN Minghui1,WU Ying1,YU Kejun1,WANG Xinrui1,YE Yaping1,QIAN Jiajun1,ZHANG Kaiyao1,WANG Siyi1,CHEN Dafu1,2,GUO Rui1,2*
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| (1. College of Animal Sciences(College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, China;2. Apitherapy Research Institute, Fujian Agriculture and Forestry University, Fuzhou 350002, China) |
| Abstract: |
| The aims of this study are to obtain the sequence of Nosema ceranae NCRBL-081 gene and predict and analyze the NCRBL-081 coding protein. NCRBL-081 gene is cloned by RT-PCR. The encoded protein of NCRBL-081 gene is predicted by related bioinformatics tools. The results indicate that: NCRBL-081 contains 630 nucleotides encoding a hydrophilic protein of 209 amino acids; NCRBL-081 encoded protein cis consisted of one signal peptide, six serine phosphorylation sites, five tyrosine phosphorylation sites and one transmembrane domain; Additionally, the encoded protein has 20. 10% of alpha helix, 8. 61% of beta fold, 25. 84% of extended chain and 45. 45% of irregular coil, and it has typical characteristics of type II ribosome inactivating protein family; Various RBL proteins from the same microsporidian species display closer evolutionary distance, while RBL proteins from different microsporidian species have more distant evolutionary distance; Moreover, NCER_100581 and AAJ76_100008081 in N. ceranae have a closer genetic relationship. The study obtained the full sequence of NCRBL-081 in N. ceranae and revealed the molecular characteristics of NCRBL-081 coding protein, which lay a foundation for the following functional analyses. |
| Key words: Nosema ceranae ricin B-lectin molecular cloning bioinformatics phylogenetic tree |
