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乳牛瘦蛋白基因荧光定量RT-PCR检测方法的建立
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张才1,牛淑玲1,夏成2,刘国文1,王哲1
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（1.吉林大学 畜牧兽医学院，吉林 长春130062;
2.黑龙江八一农垦大学 动物科技学院,黑龙江 大庆163319）
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摘要：
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根据GenBank中瘦蛋白（leptin）基因序列设计合成了引物和探针，对荧光定量PCR的方法进行了方法学的评估，建立了TaqmanMGB荧光定量RT-PCR检测方法。结果表明，由pMD-18T瘦蛋白所构建的标准曲线线性关系良好，建立的瘦蛋白基因荧光定量PCR检测方法灵敏度高、特异性强（可检测出低于10个拷贝/μL的样品）,准确可靠。 |
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关键词：
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乳牛；瘦蛋白基因；荧光定量RT-PCR |
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中图分类号：S 815.3：Q 503 文献标识码：
A 文章编号：1673-4696(2006)05-0393-03 |
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Development of a fluorescent quantitative RT-PCR assay
for detection of expression of leptin mRNA |
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ZHANG Cai1,NIU Shu-ling1,XIA Cheng2 ,LIU
Guo-wen1,WANG Zhe1 |
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(1.College of Animal Science and Veterinary Medicine , Jilin University，Changchun 130062,China; 2.College
of Animal Science and Technology, Heilongjiang August First Land Reclamation University,Daqing 163319, China) |
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Abstract: The probe and primers were designed and synthesized according to the leptin sequence avai-lable in GenBank. Then a real-time RT-PCR assay was developed and assessed. Results showed that the standard curve made by pMD-18T leptin had good linear dependence, and the fluorescent quantitative RT PCR assay was sensitive (it could detect <10 copies/μL of plasmid DNA) and specific. |
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Key words: dairy cow; leptin gene; fluorescent quantitative RT-PCR |
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