本文關(guān)鍵詞： HOXA11基因 啟動子 T載體 pEGFP-1 Ishikawa細(xì)胞　出處：《大連醫(yī)科大學(xué)》2007年碩士論文　論文類型：學(xué)位論文

【摘要】： 同源框基因(homeobox gene)是一類調(diào)控胚胎發(fā)育和細(xì)胞分化的轉(zhuǎn)錄調(diào)節(jié)基因,這類基因中都存在一個(gè)由183個(gè)堿基對(bp)組成的、高度保守的同源框(homeobox)序列。HOX基因是同源框基因中的一大類,其表達(dá)的蛋白質(zhì)是一類轉(zhuǎn)錄調(diào)節(jié)因子,都有一個(gè)由相應(yīng)同源框序列編碼的約61個(gè)氨基酸組成的同源域(homeodomain)結(jié)構(gòu),這一結(jié)構(gòu)通過與順式調(diào)控元件相結(jié)合,實(shí)現(xiàn)HOX基因?qū)Π谢虻恼{(diào)控。HOXA11屬HOX基因家族中的一員,在胚胎尾部體節(jié)中表達(dá),主要參與子宮形態(tài)的發(fā)生和發(fā)育;HOXA11基因的產(chǎn)物作為轉(zhuǎn)錄因子參與調(diào)節(jié)子宮內(nèi)膜形態(tài)發(fā)育和分化成熟。卵巢激素與HOX基因表達(dá)的關(guān)系成為近年來生殖領(lǐng)域的研究熱點(diǎn)。目前認(rèn)為HOX基因是性激素對子宮調(diào)節(jié)鏈中的重要介導(dǎo)基因,雌、孕激素對內(nèi)膜的多種調(diào)節(jié)作用主要通過HOX基因介導(dǎo)完成的。本課題組以往的研究發(fā)現(xiàn),HOXA11在子宮內(nèi)膜的表達(dá)受雌、孕激素調(diào)節(jié),在分泌中期HOXA11表達(dá)變化最明顯;并且首次發(fā)現(xiàn),分泌中期內(nèi)膜腺上皮HOXA11的表達(dá)明顯減弱甚至消失、孕激素對內(nèi)膜腺上皮HOXA11基因的表達(dá)具有負(fù)調(diào)控作用。這種負(fù)調(diào)控作用與分泌中期內(nèi)膜的分化、成熟以及子宮接受態(tài)(receptive state)的形成密切相關(guān)。但是,有關(guān)孕激素對內(nèi)膜腺上皮HOXA11基因的負(fù)調(diào)控作用機(jī)制,目前尚不清楚;HOXA11基因的啟動子區(qū)是否存在著孕激素反應(yīng)元件尚未見報(bào)道。因此,本研究的目的是:通過構(gòu)建HOXA11 pEGFP-啟動子重組質(zhì)粒,確定HOXA11基因啟動子的位置;證明HOXA11啟動子區(qū)是否存在孕激素反應(yīng)元件,為進(jìn)一步研究孕激素對內(nèi)膜腺上皮負(fù)調(diào)控機(jī)制奠定實(shí)驗(yàn)基礎(chǔ)。 主要方法: 從基因庫中查找人HOXA11基因序列,分別在正義鏈和反義鏈上預(yù)測兩段啟動子區(qū)域(分別命名為啟動子1,啟動子2),設(shè)計(jì)PCR引物;從人蛻膜組織中提取基因組DNA,用PCR方法得到克隆目的基因片段;
[Abstract]:Homeobox gene (homeobox gene) is a kind of transcriptional regulation of embryonic development and cell differentiation regulation genes, these genes are one of the 183 base pairs (BP) composed of highly conserved homeobox.HOX gene sequence (homeobox) is a large class of homeobox genes, the expression of protein is a transcription factor, a homeodomain by a corresponding homeobox encoding a sequence of about 61 amino acids (homeodomain) structure, this structure is combined with cis regulatory elements, a member of the regulation of HOX gene on.HOXA11 gene belongs to HOX gene family, expressed in embryonic tail somite, mainly involved in the occurrence and development of uterine morphology; HOXA11 gene product as a transcription factor involved in the regulation of endometrial morphological development and differentiation. The relationship between ovarian hormones and HOX gene expression has become the field of reproductive research in recent years Hotspot. Now that the HOX gene is an important regulation of sex hormones on uterine lies in the chain of gene, estrogen, progesterone on endometrial variety regulation is mainly mediated by HOX gene complete. Our group's previous study found that HOXA11 expression in endometrial estrogen and progesterone regulate in mid secretory expression of HOXA11 the most obvious change; and for the first time found that the expression of the mid secretory endometrial epithelial cells of HOXA11 decreased and even disappeared with the negative regulation of progesterone on the expression of endometrial epithelial cells. HOXA11 gene differentiation and secretion of the negative regulation of the medium term membrane, maturation and uterine receptivity (receptive state) are closely related. However, negative the regulation mechanism of progesterone on endometrial epithelial HOXA11 gene, it is unclear; the promoter region of HOXA11 gene and the existence of progesterone response element has not been reported for. The purpose of this study is to identify the location of the promoter of HOXA11 gene by constructing the recombinant plasmid of HOXA11 pEGFP- promoter, and to prove whether there is progesterone response element in HOXA11 promoter region, so as to lay an experimental foundation for further study of the negative regulation mechanism of progesterone on the endometrial glandular epithelium.
The main methods:
The sequence of human HOXA11 gene was searched from the gene pool, and the two promoter regions were predicted on the chain of justice and antisense chain, respectively, named as promoter 1, promoter 2. PCR primers were designed, genomic DNA was extracted from human decidual tissue, and the target gene fragment was obtained by PCR.

【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R346

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 羅宏志,王弘毅,王介東,王莉芬,朱正美;雌/孕激素對原代培養(yǎng)的子宮內(nèi)膜上皮細(xì)胞HOXA11和PR基因表達(dá)的影響[J];生殖與避孕;2002年01期

2 李丹,王莉芬,范成明,王化麗,姜繼勇;孕激素及間質(zhì)細(xì)胞培養(yǎng)液對原代培養(yǎng)內(nèi)膜腺上皮HOXA11基因表達(dá)的影響[J];生殖與避孕;2005年08期

3 王莉芬,王弘毅,武捷,羅宏志,朱正美,王介東;小劑量米非司酮對分泌中期人子宮內(nèi)膜HOXA11基因表達(dá)的影響[J];生殖醫(yī)學(xué)雜志;2000年03期

4 李丹,王莉芬,范成明,王乃玉,王化麗,姜繼勇;孕酮對培養(yǎng)人子宮內(nèi)膜腺上皮細(xì)胞HOXA11基因表達(dá)的影響[J];生殖醫(yī)學(xué)雜志;2005年05期

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本文編號：1497286