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成年核移植山羊生长相关基因的表达分析
引用本文:邢宝松,徐银学,成勇,刘红林,杜淼.成年核移植山羊生长相关基因的表达分析[J].遗传学报,2007,34(8):709-719.
作者姓名:邢宝松  徐银学  成勇  刘红林  杜淼
作者单位:1. 南京农业大学动物科技学院,南京,210095;河南省农业科学院畜牧兽医研究所,郑州,450002
2. 南京农业大学动物科技学院,南京,210095
3. 扬州大学兽医学院,扬州,225009
4. 中国科学院遗传与发育生物学研究所,北京,100101
摘    要:体细胞核移植过程有可能影响克隆动物生长相关基因尤其是印迹基因的表达水平。本研究运用同源引物 PCR 扩增、RACE 技术并结合同源克隆策略, 克隆了 7 个山羊生长相关基因包括 3 个印迹基因(H19、IGF2 和 IGF2R)和 4 个非印迹基因(IGF1、IGF1R、GHR 和 GHSR)的完全 CDS 或者部分 cDNA 序列, 经生物信息学技术确认后, 用荧光实时定量 PCR 对 8只成年克隆山羊中这些基因的表达水平进行分析, 结果表明 3 个印迹基因中 IGF2R 基因表达水平极显著高于对照组的自然繁殖山羊(P<0.01), 而 H19 和 IGF2 的表达则没有很大区别; 4 个非印迹基因中只有 IGF1R 的表达水平极显著高于对照组(P<0.01), IGF1、GHR 和 GHSR 的表达与对照组相似。表明即使在表型正常的成年克隆动物也存在一定的表观遗传异常。通过对获得完全 CDS 和 3′UTR 的 IGF2 基因经过生物信息学分析表明, 山羊 IGF2 基因包含一个 540 bp 的开放阅读框 (ORF)编码 179 个氨基酸。IGF2 基因 cDNA 序列和氨基酸序列以及其它基因部分序列比较分析表明, 山羊所有这些基因与绵羊的同源性要高于同牛的同源性。

关 键 词:山羊  核移植  基因表达  荧光实时定量PCR  印迹基因
收稿时间:9 November 2006
修稿时间:2006-11-09

Overexpression of IGF2R and IGF1R mRNA in SCNT-produced Goats Survived to Adulthood
Baosong Xing,Yinxue Xu,Yong Cheng,Honglin Liu,Miao Du.Overexpression of IGF2R and IGF1R mRNA in SCNT-produced Goats Survived to Adulthood[J].Journal of Genetics and Genomics,2007,34(8):709-719.
Authors:Baosong Xing  Yinxue Xu  Yong Cheng  Honglin Liu  Miao Du
Institution:1.College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China; 2. Institute of Animal Science, Henan Academy of Agricultural Science, Zhengzhou 450002, China; 3. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; 4. Institute of Genetics and Developmental Biology, Chinese Academy of Science, Beijing 100101, China
Abstract:The procedure of somatic cell nuclear transfer (SCNT) is likely to affect the expression level of growth-related genes especially imprinting genes. In this study, expressions of growth-related genes including three imprinting genes (H19, IGF2, and IGF2R) and four non-imprinting genes (IGF1, IGF1R, GHR, and GHSR) in adult nuclear transferred (NT) goats were investigated by real-time PCR. The expressions of these genes in adult clones were found largely normal, but IGF2R and IGF1R were more highly expressed in cloned goats than in non-NT goats (P < 0.01). Analysis on mono-allelic expression pattern of imprinting genes indicated that mono-allelic expression patterns of H19 and IGF2 in cloned goats were similar to that in non-NT goats. In addition, the sequence of goat IGF2 gene and the putative amino acid sequence were obtained. The 986 nucleotide cDNA of goat IGF2 gene contained an open-reading frame of 540 nucleotides coding for 179 amino acids. Both cDNA sequence and amino acid sequence of IGF2 in goat showed their higher homology with that in sheep than in cattle; the partial cDNA fragments of H19, IGF2R, GHSR, IGF1R, and GHR in goat were also cloned and sequenced, which shared higher sequence identities with those in sheep than in cattle.
Keywords:goat  nuclear transfer  gene expression  real-time PCR  imprinting gene
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