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小鼠植入前发育过程中蛋白酪氨酸磷酸酶表达的动态变化:通过实时PCR进行半定量分析

Dynamic changes in the expression of protein tyrosine phosphatases during preimplantation mouse development: semi-quantification by real-time PCR.

作者信息

Hara Kentaro T, Sakuma Yuki, Sakai Senkiti, Nagata Masao, Aoki Fugaku

机构信息

Graduate School of Frontier Sciences, University of Tokyo, Shinryoiki-Seimei Building 302, Kashiwa, Chiba 277-8562, Japan.

出版信息

J Reprod Dev. 2003 Aug;49(4):323-8. doi: 10.1262/jrd.49.323.

Abstract

Protein tyrosine phosphatase (PTP) expression was examined in preimplantation mouse embryos. We previously reported that SHP-2, LAR, PTPT9, SHP-1, and mRPTPB were expressed in preimplantation mouse embryos. Here, we examined changes in the expression levels of these PTPs during preimplantation development. cDNA was obtained by reverse transcription of embryo mRNA, amplified with 10 PCR cycles, and then subjected to real-time fluorescence-monitored PCR. Experiments with an mRNA dilution series revealed that the data obtained matched the quantities of mRNA used. The measurements obtained with real-time fluorescence-monitored PCR showed that the expression of each PTP mRNA changed dynamically, and that each had a different expression pattern. This suggests that PTPs are involved in the regulation of growth and differentiation during preimplantation development.

摘要

在植入前的小鼠胚胎中检测了蛋白酪氨酸磷酸酶(PTP)的表达。我们之前报道过,SHP-2、LAR、PTPT9、SHP-1和mRPTPB在植入前的小鼠胚胎中表达。在此,我们检测了这些PTP在植入前发育过程中表达水平的变化。通过胚胎mRNA的逆转录获得cDNA,经10个PCR循环扩增,然后进行实时荧光监测PCR。对mRNA稀释系列进行的实验表明,获得的数据与所用mRNA的量相符。实时荧光监测PCR获得的测量结果显示,每种PTP mRNA的表达动态变化,且每种都有不同的表达模式。这表明PTP参与了植入前发育过程中的生长和分化调控。

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