猪蛔虫缺氧诱导因子在雌雄成虫转录差异性的研究  

Differences in trasncription of hypoxia-inducible factor-1 subunits in male and female Ascaris suum

作　　者：宋杰[1] 何庆丰[1] 陈维春[2] 

机构地区：[1]广东医学院寄生虫学教研室,广东东莞523808 [2]广东医学院医学分子诊断重点实验室

出　　处：《中国病原生物学杂志》2015年第11期999-1001,1006,共4页Journal of Pathogen Biology

摘　　要：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。Objectives To detect differences in transcription of the hypoxia-inducible factor1 （hif-1） gene from adult male and female Ascaris suurn and to examine molecular mechanisms for sex differences in A. suum in order to provide in- formation for coatrol of ascariasis. Methods Total RNA was extracted from male and female adult A. suum and cDNA was synthesized by reverse transcription. Adult A. suum of different sexes were examined for transcription of the hif-lα and hif-1β genes using real-time PCR. The actin gene served as a control. Results The RNA obtained had a concentra- tion of 1.5-2.0 μg/lal and A260/A280 was between 1.9 and 2.0. Primers for PCR were satisfactory as indicated by a good dissociation curve and Rn vs Cycle. The 2 △△Ct method was used to analyze the expression of the genes in question. In fe- male A. suum, 2-△△Ct for transcription of hif-1α was 15. 99±3.65 and 2-△△Ct, for transcription of hif-lβ was 26.26!7. 09. In male A. suurn, 2-zxAct for transcription of hif-1α was 1.02±0.06 and 2-△△Ct, for transcription of hif-1β was 1.01 ±0.05. The difference in 2-△△Ct values was statistically significant （P〈0.05）. Conclusion There was a higher level of hif-1α and hif-1β transcription in female adult A. suum than in male A. suum, indicating that the female adult A. suum are better adapted to hypoxia in a host.

关 键 词：猪蛔虫 缺氧诱导因子 定量PCR 转录 

分 类 号：R383.11[医药卫生—医学寄生虫学]