食品科学

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金属硫蛋白基因表达水平评估青海不同地区的重金属污染程度

薛艳锋1,罗毅皓2,刘祥军2,谢凤莲2,蒋国斌2,孙万成2,*   

  1. 1.青海大学畜牧兽医科学院,青海 西宁 810016;2.青海大学农牧学院,青海 西宁 810016
  • 出版日期:2015-09-25 发布日期:2015-09-11
  • 通讯作者: 孙万成
  • 基金资助:

    国家自然科学基金重点项目(31160125)

Assessment of Heavy Metal Pollution by Expression of Yak Metallothionein Gene in Qinghai

XUE Yanfeng1, LUO Yihao2, LIU Xiangjun2, XIE Fenglian2, JIANG Guobin2, SUN Wancheng2,*   

  1. 1. Academy of Animal Science and Veterinary Medicine, Qinghai University, Xining 810016, China;
    2. College of Agriculture and Animal Husbandry, Qinghai University, Xining 810016, China
  • Online:2015-09-25 Published:2015-09-11
  • Contact: SUN Wancheng

摘要:

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cDNA,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cDNA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cDNA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。

关键词: 金属硫蛋白, 实时定量聚合酶链式反应, 基因表达, 牦牛, 酶联免疫吸附测定

Abstract:

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 cDNA 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 (mRNA level) and ELISA reaction (protein level) displayed that the most serious pollution regions of heavy
metal were Gonghe and Gangcha.

Key words: metallothionein, real-time polymerase chain reaction, gene expression, yak, enzyme-linked immuno sorbent assay

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