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相对定量法检测nestin-GFP转基因小鼠外源基因整合拷贝数 被引量：1

Relative quantification for the copies of exogenous gene in transgenic nestin-GFP mice
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摘要 目的计算出转基因小鼠nestin-GFP的外源基因整合拷贝数,为研究该模型中外源基因遗传及表达的稳定性打下基础。方法用荧光定量PCR的方法对F0代的2只小鼠基因组做检测。先根据已知拷贝数的GFP(green fluorescent protein)和GAPDH来制作标准曲线和方程,然后计算出基因组中二者所含拷贝数的比例来计算外源基因的拷贝数。结果 2只F0的外源基因拷贝数分别是3和4。结论 nestin-GFP转基因小鼠的外源基因整合拷贝数分别是3和4。 Objective To estimate the copies of exogenous gene in transgenic nestin-GFP mice for laying the foundation on the further study of the genetics and expression stability of the exogenous gene.Methods The F0 generation heterozygous transgenic netin-GFP expressing the green fluorescent protein（GFP） was used to estimate the copies of exogenous gene by quantitative real-time PCR.The standard curve and equation were made according to the known copy number GFP and internal GAPDH to calculate the foreign gene copies by calculating the two copy number ratios.Results The F0 generation heterozygous transgenic netin-GFP exogenous gene copy numbers of two mice were three and four.Conclusion The exogenous gene copy number of transgenic nestin-GFP is three and four,respectively.
作者 李振林 郑文宏 贾俊双 安靓
机构地区 南方医科大学组织学与胚胎学教研室 南方医科大学肿瘤研究所
出处 《山西医科大学学报》 CAS 2013年第1期72-75,85,86,共6页 Journal of Shanxi Medical University
基金 国家自然科学基金委 广东省政府联合重点基金资助项目(U0732006)
关键词 定量PCR 外源基因 拷贝数 quantitative real-time PCR exogenous gene copies
分类号 R-332 [医药卫生]
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参考文献7

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二级参考文献13

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共引文献27

同被引文献8

  • 1王晓建,杨旭,宋晓东,张禅那,刘继斌,邸冉,张连峰,惠汝太.实时荧光定量PCR法检测转基因小鼠拷贝数[J].中国实验动物学报,2007,15(3):170-174. 被引量：27
  • 2王育花,赵森,陈芬,肖国樱.利用实时荧光定量PCR法检测转基因水稻外源基因拷贝数的研究[J].生命科学研究,2007,11(4):301-305. 被引量：49
  • 3Ingham, D. J. , Beer, S. , Money, S. , & Hansen, G. (2001). Quantitative real - time PCR assay for determining transgene copy number in transformed plants. Biotechrdques , 31 ( 1 ) : 132 - 141.
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  • 8杨继山,董晓,潘庆杰.抗凝血酶Ⅲ转基因山羊外源基因拷贝数及其蛋白表达量的测定[J].生物技术通讯,2010,21(3):393-396. 被引量：4

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山西医科大学学报
2013年 第1期

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