中国寄生虫学与寄生虫病杂志 ›› 2005, Vol. 23 ›› Issue (5): 1-261.

• 论著 •    下一篇

斯氏按蚊转录因子Relish基因克隆、定位及不同食源条件下表达

杨松*,黄复生   

  1. 第三军医大学基础医学部病原生物学教研室,重庆 400038
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2005-10-30 发布日期:2005-10-30

Cloning, Location and Differential Analysis of Transcription FactorRelish Gene from Anopheles stephensi under Different Feeding Conditions

YANG Song*,HUANG Fu-sheng   

  1. Department of Pathogen Biology,The Third Military Medical University,Chongqing 400038,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2005-10-30 Published:2005-10-30

摘要: 目的 对斯氏按蚊转录因子Relish基因进行克隆、定位和差异分析,初步探讨转录因子Relish在前酚氧化酶(PPO)级联与疟原虫卵囊黑化中的信号调控作用。 方法 设计简并引物,对全蚊cDNA模板进行RT-PCR扩增,产物纯化、克隆、测序和鉴定,利用特异引物分别从血细胞或中肠扩增目的基因,于不同食源条件下进行半定量分析和原位核酸分子杂交定位。 结果 获得了1种斯氏按蚊Relish cDNA部分序列,与冈比亚按蚊Relish基因很相似,并存在于中肠和血细胞,半定量分析显示,感染约氏疟原虫6、12、24和48 h或吸硝喹糖水12和24 h,于诱导约氏疟原虫卵囊黑化之前表达明显增强,原位核酸分子杂交证实血细胞和中肠有表达。 结论 斯氏按蚊转录因子Relish可能在疟原虫感染或/和约氏疟原虫卵囊黑化调控中发挥重要作用。

关键词: 斯氏按蚊, 转录因子, 信号调控

Abstract: Objective To clone, locate and differentially analyze the transcription factor Relish gene from Anopheles stephensi, and to examine its signals-modulating action on prophenoloxidase cascade and melanization of Plasmodium yoelii oocysts. Methods Relish cDNA of total mosquitoes was amplified by RT-PCR with degenerated primers. Target PCR product was purified,cloned,sequenced and identified. Special Relish gene was amplified with specific primers from hemocytes or midgut,respectively. Semi-quantitative analysis was made under different feeding conditions. Relish message ribonucleic acid was identified with hybridization in situ. Results One cDNA segment of Relish similar to An.gambiae was acquired from An. stephensi. The same Relish gene was also manifested in the hemocytes and midgut. Marked up-regulation expression of Relish was observed at 6,12,24 or 48 h of Plasmodium yoelii infection and at 12 and 24 h after sucking nitroquine-acetate sucrose solution,that was before inducible oocyst melanization. Relish was also expressed in the hemocytes and midguts by ISH. Conclusion Transcription factor Relish of An. stephensi might play a role in signal modulation of Plasmodium yoelii infection and oocyst melanization.

Key words: Anopheles stephensi, Transcription factor, Signal modulation