【摘要】： 精子發(fā)生是一個(gè)多階段，多步驟持續(xù)的細(xì)胞分裂和分化過程，它受到性染色體和常染色體上一系列基因的調(diào)控。研究表明：鋅指蛋白基因家族作為哺乳動(dòng)物最大的基因家族之一，參與了精子發(fā)生和成熟的各個(gè)階段的基因表達(dá)調(diào)控。精子發(fā)生相關(guān)基因ZNF230屬于鋅指蛋白家族，位于11p15，是我室克隆的一個(gè)與人類精子發(fā)生過程相關(guān)的基因。 本研究圍繞精子發(fā)生相關(guān)基因ZNF230的功能研究，分為兩個(gè)部分。 第一部分：構(gòu)建小鼠鋅指蛋白基因Znf230條件性打靶載體，為產(chǎn)生Znf230基因條件性敲除的小鼠模型做好準(zhǔn)備工作。設(shè)計(jì)和合成特異性引物，經(jīng)PCR從小鼠的基因組中擴(kuò)增出5′同源臂、3′同源臂及錨定序列，長(zhǎng)度分別為4，4，1kb的片段，反向插入pBS載體的Neo基因的兩側(cè)，從而構(gòu)建小鼠Znf230條件性打靶載體Znf230-pBS(5)。經(jīng)過限制性內(nèi)切酶及DNA測(cè)序鑒定，證實(shí)該條件性基因打靶載體含有的同源序列與Genebank公布的基因序列一致，表明載體構(gòu)建成功。PCR技術(shù)和定向克隆技術(shù)是構(gòu)建條件基因打靶載體簡(jiǎn)單而可靠的方法。運(yùn)用該技術(shù)獲得了小鼠鋅指蛋白基因Znf230的條件性打靶載體。 第二部分：癌-睪丸抗原(Cancer-testis antigen，CTA)是目前發(fā)展腫瘤免疫治療的理想靶分子，因其在多種腫瘤組織中均有表達(dá)，但只在正常人體的睪丸組織表達(dá)，其他組織無表達(dá)。在這部分工作中我們從蛋白和mRNA水平檢測(cè)了ZNF230在不同組織類型的腫瘤中的表達(dá)情況，我們發(fā)現(xiàn)該基因在肺癌、肝癌、腎癌、食管癌、胃癌等腫瘤中有不同程度的表達(dá)，提示該基因?yàn)镃T抗原家族的新成員。
[Abstract]:Spermatogenesis is a multi-stage, multi-step process of cell division and differentiation, which is regulated by a series of genes on sex chromosome and autosomal. As one of the largest gene families in mammals, zinc finger protein gene family is involved in the regulation of gene expression in all stages of spermatogenesis and maturation. Spermatogenesis related gene ZNF230 belongs to the zinc finger protein family located at 11p15 and is a gene related to human spermatogenesis cloned in our laboratory. This study focuses on the function of spermatogenesis related gene ZNF230, which is divided into two parts. Part one: construct the Znf230 conditional targeting vector of mouse zinc finger protein gene, and prepare the mouse model for producing Znf230 gene conditional knockout. A specific primer was designed and synthesized. The 5'homologous arm 3'homologous arm and its anchoring sequence were amplified from mouse genome by PCR. The fragments with the length of 41kb were inserted into the two sides of the Neo gene of the pBS vector. The conditional targeting vector Znf230-pBS (5) of mouse Znf230 was constructed. By restriction endonuclease and DNA sequencing, it was confirmed that the homologous sequence contained in the conditional gene targeting vector was the same as that published by Genebank. The results showed that the successful construction of vector. PCR and directional cloning were simple and reliable methods to construct conditional gene targeting vector. The conditional targeting vector of mouse zinc finger protein gene Znf230 was obtained by this technique. The second part: carcinomatous testis antigen (Cancer-testis antigen,CTA) is an ideal target molecule for the development of tumor immunotherapy, because it is expressed in many kinds of tumor tissues, but only in normal human testis, but not in other tissues. In this part of our work, we detected the expression of ZNF230 in different tissue types of tumors at the protein and mRNA levels, and we found that the gene was expressed to different degrees in lung cancer, liver cancer, renal cancer, esophageal cancer, gastric cancer, and so on. It is suggested that this gene is a new member of the CT antigen family.
【學(xué)位授予單位】：四川大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R346;R73-3

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相關(guān)期刊論文 前1條

1 楊曉,黃培堂,黃翠芬;在小鼠進(jìn)行基因打靶的研究進(jìn)展[J];科學(xué)通報(bào);2000年15期

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