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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。
Objective To study the changes of mRNA expression of nitric oxide synthase (NOS) in snail gonads at different temperatures. Methods Snails into the incubator at different temperatures (0 ℃, 15 ℃, 25 ℃) for 1 month. The total RNA was extracted by total RNA extraction kit, and the degenerate primers of NOS were designed according to the conserved sequence of mammalian NOS. Reverse transcription polymerase chain reaction (RT-PCR) was used to determine the expression of NOS mRNA in snail gonads of each experimental group. Results The bands of specific PCR products appeared in each experimental group. The expression of NOS mRNA in 0 ℃ group and 25 ℃ group was significantly higher than that in control group (P <0.01), while there was no significant difference between 15 ℃ group and control group P> 0.05). Conclusion Primers designed in this study are feasible, and temperature changes can affect the expression of NOS mRNA in Oncomelania snail.