本文選題：顯微注射 + CEA　； 參考：《鄭州大學(xué)》2006年碩士論文

【摘要】：轉(zhuǎn)基因動物技術(shù)的出現(xiàn)是20世紀(jì)生物技術(shù)領(lǐng)域中的重大事件，而生物技術(shù)將是21世紀(jì)的支柱產(chǎn)業(yè)之一，是經(jīng)濟和科技發(fā)展的關(guān)鍵技術(shù)。轉(zhuǎn)基因技術(shù)使人們能從整體、器官、細胞、分子多層次，從發(fā)育時間和組織生理空間多角度，立體多維地研究基因，并可按人類的意愿改造和修飾物種。 癌胚抗原(carcinoembryonic antigen，CEA)是胚胎發(fā)展中產(chǎn)生的抗原之一，是一種最常見的腫瘤相關(guān)抗原，是目前國際上公認(rèn)的腫瘤標(biāo)記物。其基因定位于人類19號染色體長臂(19q13.1～13.2)，分子量約為180kd，為免疫球蛋白超家族的成員。CEA在正常消化道黏膜僅少量分泌，而90％的消化道惡性腫瘤、胰腺癌，，70％非小細胞肺癌和50％乳腺癌有CEA的高表達，故CEA可作為這些腫瘤檢測和治療的靶分子。 本實驗以小鼠為研究對象，用顯微注射法將包含小鼠乳清酸蛋白基因啟動子(約2.4kb)、人癌胚抗原的編碼序列(約2.1kb)及polyA信號的轉(zhuǎn)基因構(gòu)件轉(zhuǎn)入原核期小鼠胚胎，制備了CEA轉(zhuǎn)基因小鼠，旨在研究CEA與乳腺癌等腫瘤發(fā)生、發(fā)展的關(guān)系，為以后制作小鼠乳腺生物反應(yīng)器提供經(jīng)驗和參考。主要進行了以下工作： 提純pWA-CEA質(zhì)粒，分別進行HindⅢ單酶切、HindⅢ和Xho Ⅰ雙酶切、SacⅡ和KpnⅠ雙酶切鑒定。對構(gòu)建成功的pWA-CEA質(zhì)粒用SacⅡ和KpnⅠ雙酶切得到目的基因(約4.9kb)，膠回收并純化目的片斷，溶于TE(10mmol／L Tris-HCl，pH7.4；0.2mmol／L EDTA)中，注射用DNA濃度為3μg／ml，儲存-20℃?zhèn)滹@微注射用。將雌鼠做超排卵處理，與正常雄鼠交配，取其受精卵，培養(yǎng)至單細胞雙原
[Abstract]:The emergence of transgenic animal technology is an important event in the field of biotechnology in the 20th century, and biotechnology will be one of the pillar industries in the 21st century and the key technology for the development of economy and science and technology. Transgenic technology enables people to study genes in a three-dimensional and multidimensional manner from the perspectives of whole, organ, cell and molecular levels, development time and tissue physiological space, and can modify and modify species according to human wishes. Carcinoembryonic antigen (CEA) is one of the antigens produced in embryonic development. It is one of the most common tumor-associated antigens and is an internationally recognized tumor marker. Its molecular weight is about 180 kd.The gene is located in human chromosome 19, 19q13.1 and 13.2kd. it is a member of the immunoglobulin superfamily, which secretes only a small amount of CEA in the normal gastrointestinal mucosa, while 90% of the malignant tumors of the digestive tract. 70% of non-small cell lung cancer and 50% of breast cancer have high expression of CEA, so CEA can be used as a target molecule for the detection and treatment of these tumors. In this study, mice were studied. Transgenic components containing mouse whey acid protein gene promoter (about 2.4 kb), human carcinoembryonic antigen coding sequence (about 2.1 kb) and polyA signal were transferred into prokaryotic mouse embryos by microinjection. CEA transgenic mice were prepared to study the relationship between CEA and carcinogenesis and development of breast cancer, and to provide experience and reference for the production of mouse mammary gland bioreactor in the future. The main tasks were as follows: PWA-CEA plasmids were purified and identified by single enzyme digestion of Hind 鈪

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