中国血吸虫病防治杂志 ›› 2020, Vol. 32 ›› Issue (6): 584-.

• 论著 • 上一篇    下一篇

大劣按蚊Torso-like基因鉴定 分子结构及表达特征

朱凌倩1,胡小康1,许静1,李石柱1, 2*,冯欣宇1, 2*   

  1. 1中国疾病预防控制中心寄生虫病预防控制所、国家热带病研究中心、WHO热带病合作中心、科技部国家级热带病国际联合研究中心、国家卫生健康委员会寄生虫病原与媒介生物学重点实验室(上海 200025);2上海交通大学医学院全球健康学院
  • 出版日期:2020-12-08 发布日期:2020-12-08
  • 作者简介:朱凌倩,女,硕士研究生。研究方向:媒传疾病分子生物学
  • 基金资助:
    国家自然科学基金青年科学基金(81802039);国家科技重大专项(2016ZX10004222?004)

Identification, molecular structure and expression characteristics of Torso?like gene in Anopheles dirus

ZHU Ling-Qian1, HU Xiao-Kang1, XU Jing1, LI Shi-Zhu1, 2*, FENG Xin-Yu1, 2*   

  1. 1 National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Chinese Center for Tropical Diseases Research, WHO Collaborating Centre for Tropical Diseases, National Center for International Research on Tropical Diseases, Ministry of Science and Technology, National Health Commission Key Laboratory of Parasites and Vector Biology, Shanghai 200025, China; 2 One Health Center, Shanghai Jiaotong University School of Medicine, China
  • Online:2020-12-08 Published:2020-12-08

摘要: 目的 鉴定大劣按蚊Torso?like(tsl)基因及表达特征,为深入研究tsl基因功能提供理论基础。方法 根据黑腹果蝇和冈比亚按蚊tsl基因,检索大劣按蚊全基因组并鉴别大劣按蚊tsl基因,设计特异性引物,并采用PCR和逆转录PCR技术扩增目的基因。利用生物信息学软件分析tsl基因编码TSL蛋白的理化性质、信号肽、跨膜结构、蛋白质二级结构和三级结构,并进行系统发育进化分析。利用实时荧光定量PCR技术检测该基因在大劣按蚊各组织中的表达水平。结果 大劣按蚊tsl基因全长16 751 bp,CDS区长1 134 bp,编码377个氨基酸。TSL蛋白属于亲水性稳定蛋白。经生物信息学预测,TSL蛋白为位于膜外的分泌蛋白,且包含信号肽;其二级结构含α?螺旋(51.72%)、延伸链(12.20%)、β?折叠(4.78%)和无规则卷曲(31.30%),且能以5cj9.1.A为模板进行3D同源建模。系统发育进化分析发现大劣按蚊TSL蛋白与法老按蚊TSL蛋白亲缘关系较近。实时荧光定量PCR检测显示,tsl基因在大劣按蚊头、胸、腹、足均有表达,且在头部表达量最高、足部表达量较低。结论 本研究从基因组水平鉴定了大劣按蚊tsl基因,分析了其蛋白结构及组织特异性表达特征,为进一步深入研究该基因功能奠定了基础。

关键词: 大劣按蚊, Torso?like基因, 鉴定, 分子结构, 生物信息学

Abstract: Objective To characterize Torso?like (tsl) gene and investigate its expression characteristics in Anopheles dirus, so as to provide a theoretical basis for subsequent functional studies of the tsl gene. Methods According to the coding sequences of Drosophila melanogaster and An. gambiae tsl genes, the complete genome of An. dirus was retrieved and the An. dirus tsl gene was characterized. Specific primers were designed and the target gene was amplified using PCR and reverse?transcription PCR assays. The physicochemical properties, signal peptide, transmembrane structure, secondary structure and tertiary structure of the encoded protein TSL were analyzed using bioinformatics tools, and a phylogenetic analysis was performed. In addition, the specific expression of the tls gene was detected in various tissues of An. dirus using a quantitative real?time PCR assay. Results  The An. dirus tsl gene was 16 751 bp in length with a CDS region of 1 134 bp, encoding 377 amino acids, and the encoded TSL protein was a stably hydrophilic protein. The TSL protein was predicted to be a secretory protein that was located in extra?membrane regions containing signal peptides. The secondary structure of the TSL protein contained α?helix (51.72%), extended strand (12.20%), β?bridge (4.78%) and random coil (31.30%) in the secondary structure, and a 3D homology model was generated using 5cj9.1.A as a template. Phylogenetic analysis revealed a close genetic relationship in the TSL protein between An. dirus and An. farauti. In addition, quantitative real?time PCR assay detected the tsl gene expression in the head, chest, abdomen and foot of An. dirus, with the highest expression in the head and low expression in the foot. Conclusion The tsl gene is characterized in An. dirus at a genomic level, and the prediction of the TSL protein structure and the elucidation of the tissue?specific tsl gene expression in An. dirus provide a basis for the further studies on the gene functions.

Key words: Anopheles dirus, Torso?like gene, Characterization, Molecular structure, Bioinformatics analysis

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