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目的检测雌、雄成虫猪蛔虫缺氧诱导因子转录水平的差异性,探讨猪蛔虫性别差异的分子机制,为蛔虫病的防治提供参考。方法提取猪蛔虫雌、雄成虫总RNA,反转录合成cDNA,以蛔虫actin基因为内参,采用实时荧光定量PCR检测hif-1α和hif-1β基因的转录水平。结果提取的RNA浓度为1.5-2.0μg/μl,A260/A280位于1.9-2.0之间。实时荧光定量PCR显示引物具有良好的溶解曲线和扩增曲线,检测雌成虫hif-1α和hif-1β基因表达的2-△△Ct分别为15.99±3.65和26.26±7.09,雄虫分别为1.02±0.06和1.01±0.05,差异均有统计学意义(P<0.05)。结论猪蛔虫hif-1α和hif-1β基因转录水平雌成虫高于雄成虫,表明雌虫更能适应宿主体内低氧环境。
Objective To detect the difference of transcriptional level of hypoxia inducible factor between Asiogenia and Asiatic male and female adults and explore the molecular mechanism of Ascaris suum sex differences so as to provide reference for the prevention and treatment of Ascariasis. Methods The total RNA was extracted from female and male adult Ascaris suum inoculation and the cDNA was reverse transcribed. The actin gene of Ascaris was used as an internal control, and the transcriptional levels of hif-1α and hif-1β were detected by real-time fluorescence quantitative PCR. Results The RNA concentration was 1.5-2.0 μg / μl and A260 / A280 was between 1.9-2.0. Real-time fluorescence quantitative PCR showed that the primers had good dissolution curves and amplification curves. The 2 -ΔΔCt for detecting the expression of hif-1α and hif-1β genes in female adults was 15.99 ± 3.65 and 26.26 ± 7.09, respectively, while the male ones were 1.02 ± 0.06 and 1.01 ± 0.05, the differences were statistically significant (P <0.05). Conclusion The female adults of hirc-1α and hif-1β gene transcriptional levels of Ascaris suum than those of adult male show that the female can adapt more to the hypoxic environment in the host.