| 马氏珠母贝外套膜中央膜与边缘膜荧光定量PCR分析中内参基因的筛选 |
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| 引用本文： | 闫芳,罗少杰,田群莉,焦钰,邓岳文.马氏珠母贝外套膜中央膜与边缘膜荧光定量PCR分析中内参基因的筛选[J].湛江海洋大学学报,2014(6):6-11. |
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| 作者姓名： | 闫芳 罗少杰 田群莉 焦钰 邓岳文 |
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| 作者单位： | 广东海洋大学水产学院,广东湛江524025 |
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| 基金项目： | 国家自然科学基因（31272635,31201023,41206141）;广东海洋大学博士启动项目（1212318） |
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| 摘 要： | 为筛选能在马氏珠母贝外套膜边缘膜与中央膜区域稳定表达的内参基因,应用荧光定量PCR 技术,对3-磷酸脱氢酶（GAPDH）、β-肌动蛋白（β-actin）、胆色素原脱氨酶（PBGD）、微管蛋白（TUB）、TATA 盒结合蛋白（TBP）、磷脂酶A2（PLA-2）、葡萄糖苷酶（GUSB）、18S 核糖体rRNA（18SrRNA）等8 个常用的内参基因在马氏珠母贝外套膜边缘膜与中央膜区域的表达情况进行分析,并利用geNorm、NormFinder、BestKeeper 及RefFinder 软件对8 个内参基因的表达稳定性进行分析.结果表明：8 个内参基因均可获得特异性扩增产物,但稳定性各异, 其在外套膜边缘膜和中央膜区的表达稳定性顺序依次为PBGD/TBP〉TUB〉GUSB〉18SrRNA〉PLA-2〉GAPDH〉 β-actin;在荧光定量PCR 中,PBGD 和TBP 在马氏珠母贝外套膜边缘膜和中央膜区的表达比较稳定,为研究贝壳矿化机制中理想的内参基因.
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| 关 键 词： | 荧光定量PCR 边缘膜 中央膜 内参基因 马氏珠母贝 |
Screening of Reference Genes for Real-time PCR in Mantle Central andMantle Edge from Pinctada martensii |
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| YAN Fang,LUO Shao-jie,TIAN Qun-li,JIAO Yu,DENG Yue-wen.Screening of Reference Genes for Real-time PCR in Mantle Central andMantle Edge from Pinctada martensii[J].Journal of Zhanjiang Ocean University,2014(6):6-11. |
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| Authors: | YAN Fang LUO Shao-jie TIAN Qun-li JIAO Yu DENG Yue-wen |
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| Institution: | (Fisheries College of Guangdong Ocean University,Zhanjiang 524025,China) |
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| Abstract: | The mineralized layer of the shell from Pinctada martensii is divided into prismatic layer andnacreous layer. It is generally acknowledged that the prismatic layer and nacre is formed by the mantleedge and mantle central, respectively. To obtain the most stable reference gene between mantle edge andmantle central, quantitative PCR was adopted to analyze eight traditional reference genes,including-3-phosphate dehydrogenase （GAPDH）, β-actin （β-actin）, porphobilinogen deaminase （PBGD）,tubulin （TUB）, TATA box binding protein （TBP）, phospholipase A2 （PLA-2）, glucosidase （GUSB）, and18S ribosomal rRNA （18SrRNA）. Combining geNorm, NormFinder, BestKeeper and RefFindersoftware, the expression stability of eight reference genes were analyzed. The results showed specificamplification products could be obtained by the eight reference genes, but their stability was differentThe stability between mantle edge and mantle central descented as follows： PBGD/TBP〉TUB〉GUSB〉18SrRNA〉PLA-2〉GAPDH〉β-actin. These results showed the most stable reference genes betweenmantle edges and mantle central were PBGD and TBP. |
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| Keywords: | Real-time quantitative PCR mantle edge mantle central reference gene Pinctada mart ensii |
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