单个植入前胚胎mRNA差异显示方法的建立
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国家重点基础研究发展规划项目(973)(2000016107).


Establishment of Single Preimplantation Embryos Differential Display Polymerase Chain Reaction
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This work was supported by a grant from The Special Funds for Major State Basic Research of China (2000016107).

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    摘要:

    在已有的研究基础上,优化和建立了单个植入前胚胎的mRNA差异显示(single preimplantation embryo differential display polymerase chain raction,SPEDDRT-PCR).以小鼠单个成熟的卵母细胞和单个的2细胞期胚胎作为起始材料,比较二者之间基因的表达差异.选择在卵母细胞中不表达而在2细胞期表达的差异片段.进行GenBank检索和表达序列标签(EST)库电子克隆.与多胚研究方法一样,由线粒体编码的和2细胞特异表达的NADH脱氢酶亚单位2和ATPase 6证实了SPEDDRT-PCR方法是可行而且可靠的,是一种需要材料极少、应用广泛而有效的基因分离方法.

    Abstract:

    Based on previous studies, mRNA differential display using a single mouse oocyte or a 2-cell embryo as starting materials was established. cDNA fragments, which were expressed in 2-cell stage mouse embryos but not in MⅡ stage, were performed computer cloning and GenBank search. Two genes, NADH dehydrogenase gene subunit-2 and ATPase-6,which encoded by mitochondrial genome, were found to be stage-specifical expressed in 2-cell-stage pre-implantation embryos. These results, which were the same as multi-embryonic mRNA differential display, strongly suggest that single preimplantation embryos differential display polymerase chain reaction (SPEDDRT-PCR) is a credible, practical and effective method that needs only a single embryo.

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郁卫东,杨立新,李文雍,刘桂生,陈清轩.单个植入前胚胎mRNA差异显示方法的建立[J].生物化学与生物物理进展,2003,30(2):308-313

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  • 收稿日期:2002-10-17
  • 最后修改日期:2002-11-26
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