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[利用抑制性消减杂交技术鉴定着床窗期子宫内膜差异表达基因]

[Identification of differentially expressed genes in endometrium during the window of implantation using suppression substractive hybridization].

作者信息

DU Guo-ping, Zhang Wei, Wang Li, Liu Yin-kun, Zhou Jian-ping

机构信息

Department of Reproductive Endocrinology, Obstetrics and Gynecology Hospital, Fudan University, Shanghai 200011, China.

出版信息

Zhonghua Fu Chan Ke Za Zhi. 2007 Mar;42(3):187-91.

Abstract

OBJECTIVE

To identify genes differentially expressed in the window of implantation and explore the molecular basis of the development of endometrial receptivity.

METHODS

A subtracted cDNA library of the window of implantation was constructed by suppression subtractive hybridization (SSH) method. The screened clones of the subtracted library were sequenced and GenBank homology search was performed. The differential expression of ribosomal protein (RP) L7, RPL7 pseudogene (RPL7p), RPL19 and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta polypeptide (YWHAZ) were further confirmed by RT-PCR.

RESULTS

After sequencing and GenBank homology search of 50 clones, 35 differentially expressed genes were detected in the window of implantation, of which 23 were known genes, and 12 were unknown genes. Some of the known genes have been proved to be associated with implantation, while others were firstly screened out by us. The results of RT-PCR confirmed that RPL7, RPL7p, RPL19 and YWHAZ were highly expressed in the window of implantation, 0.75 +/- 0.21, 1.72 +/- 0.30, 1.23 +/- 0.31, and 1.28 +/- 0.08, respectively.

CONCLUSIONS

SSH is a useful technique to detect differential expression genes and an effective method to clone novel genes. It provides a new method to investigate the molecular basis of the development of endometrial receptivity.

摘要

目的

鉴定在着床窗口期差异表达的基因,探讨子宫内膜容受性形成的分子基础。

方法

采用抑制性消减杂交(SSH)法构建着床窗口期消减cDNA文库。对筛选出的消减文库克隆进行测序,并进行GenBank同源性搜索。通过RT-PCR进一步证实核糖体蛋白(RP)L7、RPL7假基因(RPL7p)、RPL19和酪氨酸3-单加氧酶/色氨酸5-单加氧酶激活蛋白ζ多肽(YWHAZ)的差异表达。

结果

对50个克隆进行测序和GenBank同源性搜索后,在着床窗口期检测到35个差异表达基因,其中23个为已知基因,12个为未知基因。部分已知基因已被证明与着床相关,其他则是我们首次筛选出的。RT-PCR结果证实,RPL7、RPL7p、RPL19和YWHAZ在着床窗口期高表达,分别为0.75±0.21、1.72±0.30、1.23±0.31和1.28±0.08。

结论

SSH是检测差异表达基因的有用技术,也是克隆新基因的有效方法。它为研究子宫内膜容受性形成的分子基础提供了一种新方法。

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