为揭示青海不同地区重金属污染的潜在生态风险,从玉树、果洛、海晏、刚察、共和5个地区的牦牛肝脏样品中提取总RNA,在逆转录聚合酶的作用下生成c DNA,以逆转录产物为模板,利用实时定量PCR检测牦牛金属硫蛋白基因的表达水平。若将刚察地区牦牛肝脏金属硫蛋白c DNA实时荧光定量值作为对照组,2-ΔΔCt值设定为1时,则玉树、果洛、海晏、共和地区的牦牛肝脏金属硫蛋白c DNA实时荧光定量值分别为0.669、0.624、0.529、1.320,并且通过酶联免疫反应测定的牦牛体内金属硫蛋白的含量,相互印证,从而更加准确地评估青海不同地区牦牛机体重金属暴露水平,以便对牦牛产品安全进行评价。基因表达和酶联免疫反应的实验结果都表明,共和与刚察地区的重金属污染可能较为严重。
In order to study the potential risk of heavy metal contamination in Qinghai, total RNA was extracted from yak liver samples from five regions, Yushu, Guoluo, Haiyan, Gangcha and Gonghe, and then c DNA was generated under the action of reverse transcription polymerase. The product of reverse transcription reaction was finally used to carry out RTPCR and detect the expression levels of metallothionein(MT) gene. The results showed that when yak liver sample from Gangcha was taken as the control group and the value of 2-ΔΔCt was set to be 1, the fluorescence quantitative values of yak livers from Yushu, Guoluo, Haiyan and Gonghe were 0.669, 0.624, 0.529 and 1.320, respectively. Meanwhile, these results were confirmed by comparison with those of ELISA reaction. The exposure levels of heavy metal in yak body from different regions in Qinghai were evaluated, so that we could come up with assessment to yak production safety. Both the results of gene expression(m RNA level) and ELISA reaction(protein level) displayed that the most serious pollution regions of heavy metal were Gonghe and Gangcha.