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应用双重PCR技术鉴定猪胎儿性别及其细胞系建立
引用本文:尹智,孔庆然,格日乐其木格,王振坤,张力,刘忠华.应用双重PCR技术鉴定猪胎儿性别及其细胞系建立[J].东北农业大学学报,2008,39(9).
作者姓名:尹智  孔庆然  格日乐其木格  王振坤  张力  刘忠华
作者单位:东北农业大学生命科学学院,哈尔滨,150030
摘    要:sry和zfy/x是与哺乳动物性别决定有关的重要基因。本试验针对猪sry基因设计了两对PCR引物,以zfy/x为内参基因,利用双重PCR技术对6只30d左右猪胎儿的性别进行鉴定。结果表明,所有的6个样品都有447bp(雄性)或445bp(雌性)的zfy或zfx序列的扩增带,而采用两对不同的引物对sry序列扩增后,有1个样品出现241bp和166bp的扩增带为雄性,其余的无特异性扩增带为雌性,然后采用胶原酶消化培养法建立已知性别的猪胎儿成纤维细胞系。该方法具有简单、快速、准确的特点,为转基因克隆动物研究中尽早转染目的基因以及培养特定性别的胎儿成纤维细胞系提供了可靠的依据。

关 键 词:猪胎儿  双重PCR  sry  性别鉴定

Sex identification of porcine fetus by duplex-PCR method
YIN Zhi,KONG Qingran,Gerelchimeg,WANG Zhenkun,ZHANG Li,LIU Zhonghua.Sex identification of porcine fetus by duplex-PCR method[J].Journal of Northeast Agricultural University,2008,39(9).
Authors:YIN Zhi  KONG Qingran  Gerelchimeg  WANG Zhenkun  ZHANG Li  LIU Zhonghua
Institution:YIN Zhi,KONG Qingran,Gerelchimeg,WANG Zhenkun,ZHANG Li,LIU Zhonghua(College of Life Sciences,Northeast Agricultural University,Harbin 150030,China)
Abstract:In mammals, sry and zfy/x are important to sex determination. Two pairs of PCR primers were designed and the amplification product of the zfy/x genes was used as a positive control for PCR to identify the sex of porcine fetuses by duplex-PCR method. The results showed that zfy/x genes as positive controls had the amplified 447/445 bp band in all samples, 1 of 6 samples amplified 241 bp and 166 bp bands were males and the others that didn't have amplified sequences were females, then porcine fetal fibroblast...
Keywords:sry
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