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.
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