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利用RT-PCR和3′-RACE,从中华卵索线虫Ovomermis sinensis中获得tra-1基因cDNA部分序列,该序列长800bp,共编码195个氨基酸,与GLI转录因子家族其它成员在锌指区域具有较高的相似性(70%～77%)。实时荧光定量检测显示:雌成虫tra-1相对表达量最高,与其它虫期差异极显著(P<0.01),在怀卵雌虫和胚胎发育期卵中,tra-1相对表达量明显下降,但仍显著高于其它试验组(P<0.05);其它试验组,即感染期幼虫、1～6天寄生期幼虫、寄生后期幼虫与雄成虫之间表达量无明显差异(P>0.05)。线虫在宿主体内不同的感染强度(40/1与10/1)对tra-1基因转录活性无显著影响(P>0.05)。表明tra-1基因参与调控中华卵索线虫雌性生殖系统的发育、卵的形成以及胚胎的发育。
The partial cDNA sequence of tra-1 gene was obtained from Ovomermis sinensis by RT-PCR and 3’-RACE. The cDNA sequence was 800 bp in length and encodes a protein of 195 amino acids. The sequence of tra-1 gene shared with other members of the GLI transcription factor family in the zinc finger region High similarity (70% ~ 77%). Real-time quantitative PCR assay showed that the relative expression level of tra-1 was the highest in female adults, and the difference was significant compared with other stages (P <0.01). The relative expression of tra-1 in pregnant females and embryos during embryo development decreased significantly, (P <0.05). However, there was no significant difference (P> 0.05) between the other experimental groups, that is, larvae infected during infection, larvae parasitized for 1 to 6 days, parasitized larvae and male adults. The different infection intensities (40/1 and 10/1) of nematodes in the host had no significant effect on the transcriptional activity of tra-1 (P> 0.05). The results indicated that tra-1 gene is involved in the regulation of female reproductive system development, egg formation and embryo development.