旋毛虫p46000抗原基因的转录及表达特性鉴定

Stage specific transcription and expression of p46000 antigen gene in Trichinella spiralis

根据旋毛虫p46000抗原基因和葡萄糖3-磷酸甘油醛脱氢酶(GAPDH)基因序列设计特异引物,以不同发育时期旋毛虫总RNA为模板进行SYBRGreen染料法实时定量RT-PCR检测p46000基因转录情况。制作不同发育时期旋毛虫虫体及感染组织石蜡切片,采用免疫荧光染色技术检测p46000抗原的表达及定位情况。结果显示,p46000抗原基因在旋毛虫不同发育时期均有转录,但转录水平有明显差异,转录高峰集中于成虫和肌幼虫期,新生幼虫期明显低于其他时期。天然p46000抗原蛋白定位于杆状体和虫体表面,表达高峰集中于成虫、肠道期及成囊期肌幼虫。表明,p46000抗原基因的转录与表达具有一定的时空特异性,该基因在旋毛虫的寄生过程中尤其在早期感染中具有重要作用且可能与旋毛虫的免疫逃避有关。

Specific primers were designed for p46000 antigen gene and glyceraldehyde-3-phosphate dehydrogenase（GAPDH） gene of Trichinella spiralis and the real-time quantitative RT-PCR with SYBR Green dye was performed to analyze p46000 gene expression profiles at different developmental stages of T. spiralis. Paraffin sections from parasites and infected tissues were prepared and immunofluorescence was utilized to localize p46000 gene product. The results showed that p46000 gene was transcripted at all stages of T. spiralis life cycle, but the peak levels were detected at muscle larvae and adults stages. Although native p46000 was detected in stichosome and on cuticle of the worms during the life cycle of T. spiralis, the most abundant expression of p46000 was observed at adult worms,intestinal L1 and post-encapsule L1 larvae,indicating that the transcription and expression of p46000 gene is temporal and spatial. The antigen could play an important role during the parasitism processes especially in early infection of T. spiralis. These findings provided the possibility that it could associate with immune evasion of T. spiralis.