【摘要】 目的 研究不同温度下钉螺生殖腺一氧化氮合酶(NOS)mRNA表达的变化规律。方法 钉螺放入不同温度(0℃、15℃、25℃)的培养箱内饲养1个月。用总RNA抽提试剂盒抽提总RNA,参照哺乳动物NOS的保守序列,设计出钉螺NOS的简并引物。用逆转录聚合酶链反应(RT-PCR)测定各实验组钉螺生殖腺NOS mRNA的表达量。结果 各实验组均有特异性PCR产物条带出现,0℃组和25℃组的NOS mRNA表达量明显高于对照组(P<0.01),15℃组与对照组相比差异无显著性(P>0.05)。结论 本研究设计的引物是可行的,温度变化可影响钉螺生殖腺NOS mRNA的表达。更多还原

【Abstract】 Objective To study the expression of nitric oxide synthase (NOS)mRNA in the gonad of Oncamelania hu-pensis in different temperature. Methods The snails were cultured at temperature of 0℃ , 15 ℃and 25℃for 1 month. The total RNA of each group was extracted with the RNA extraction kit. A pair of degenerate primers was designed from conserved regions of mammalian NOSs, and the expression of NOS mRNA in the gonad was measured by means of reverse transcription polymerase chain reaction (RT-PCR). Results The target genes of NOS were detected in the gonad of snails. The level of expression of snail NOS mRNA in 25℃ group and 0℃group was significantly higher than that in the control group( P <0.01), there was no significant difference for the expression products between 15℃group and control group (P>0.05). Conclusion The designation of primers of the snails was validated. The impact of temperature on the expression of snail NOS mRNA was determined, which suggested that NO plays an important role in the physiological and pathological modulation.更多还原