目的对大劣按蚊丝氨酸蛋白酶3(AdSP3)的表达进行组织定位和定量研究。方法首先利用RT-PCR方法对大劣按蚊血淋巴细胞、唾液腺和蚊胃进行AdSP3的扩增,然后利用Realtime PCR对大劣按蚊感染约氏疟原虫后不同时相点的血淋巴细胞AdSP3表达水平进行定量研究。结果从血淋巴细胞和唾液腺均成功扩增出目的片段;Reatime PCR的结果显示,吸血后24 h,感染组AdSP3在血淋巴细胞的转录水平显著高于正常对照组,吸血后5 d是AdSP3的表达高峰。结论大劣按蚊血淋巴细胞和唾液腺可表达AdSP3,AdSP3可能参与了大劣按蚊抗约氏疟原虫感染的入侵和黑化反应。
Objective To locate the tissue of Anopheles dirus where serine protease 3(AdSP3)is expressed and to quantify the expression level after infection.Methods RT-PCR was used to amplify AdSP3 from hemolymphocytes,salivary glands and midguts.The transcription level of AdSP3 in hemolymphocytes at different time points post infection was tested by Realtime PCR.Results The interesting fragments were successfully amplified from hemolymphocytes and salivary glands.The transcription of AdSP3 was significantly up-regulated by infection with Plasmodium yoelii at 24h post infection.And the expression level reached to the peak at 5d post blood meal.Conclusion Hemolymphocytes and salivary glands can express AdSP3.AdSP3 was probably involved in the mechanism of anti Plasmodium yoelii invasion and melanization.