本文選題：辜丸內(nèi)注射 + 體內(nèi)電穿孔��； 參考：《第一軍醫(yī)大學(xué)》2005年碩士論文

【摘要】：轉(zhuǎn)基因動(dòng)物在醫(yī)學(xué)、生命科學(xué)、生物制藥等領(lǐng)域有著廣泛的研究和應(yīng)用前景,因此科學(xué)家們一直在探索一種更簡(jiǎn)便易行、經(jīng)濟(jì)實(shí)用而又高效的轉(zhuǎn)基因方法。近幾年來(lái),隨著科學(xué)技術(shù)的發(fā)展,體內(nèi)基因轉(zhuǎn)移已成為用于基因治療、生物學(xué)分析等領(lǐng)域非常受歡迎的一種技術(shù),體內(nèi)電穿孔技術(shù)就是其中之一。 精原干細(xì)胞是精子的前體細(xì)胞,是雄性動(dòng)物中唯一可以自我增殖、更新并向下一代傳遞遺傳信息的細(xì)胞類型。對(duì)精原干細(xì)胞的操作和修飾在建立轉(zhuǎn)基因動(dòng)物模型、制備乳腺生物反應(yīng)器等方面有重要意義。為了探討將外源基因注射到在體睪丸生精小管內(nèi)并進(jìn)行體內(nèi)電穿孔以建立轉(zhuǎn)基因動(dòng)物的可行性,我們首先設(shè)計(jì)了低壓(30～110V/cm)和高壓(500～900V/cm)兩組不同的體內(nèi)電穿孔參數(shù)對(duì)SPF級(jí)KM雄鼠睪丸進(jìn)行在體電穿孔,以觀察不同的電穿孔條件對(duì)SPF級(jí)KM雄鼠睪丸損傷程度和生殖能力的影響,從而篩選出合適的體內(nèi)電穿孔參數(shù)。并在此基礎(chǔ)上,我們對(duì)插入有增強(qiáng)型綠色熒光蛋白(EGFP)基因的真核表達(dá)質(zhì)粒pCE-29DNA進(jìn)行2種不同的處理:(1) pCE-29DNA+轉(zhuǎn)染試劑(in vivo jetPI~(TM)+臺(tái)盼藍(lán)(TB);(2) pCE-29DNA+臺(tái)盼藍(lán)(TB)。然后將這兩種不同
[Abstract]:Transgenic animals have a wide range of research and application prospects in medicine, life science, biopharmaceutical and other fields, so scientists have been exploring a more simple, practical and efficient transgenic method. In recent years, with the development of science and technology, gene transfer in vivo has become a very popular technology in gene therapy, biological analysis and other fields, one of which is in vivo electroporation. Spermatogonial stem cells are the progenitor of spermatozoa and the only cell type in male animals that can self-proliferate, update and transmit genetic information to the next generation. The operation and modification of spermatogonial stem cells are of great significance in the establishment of transgenic animal models and the preparation of mammary gland bioreactor. In order to explore the feasibility of injecting exogenous gene into testicular seminiferous tubules in vivo and electroporation in vivo to establish transgenic animals, We first designed in vivo electroporation of the testis of SPF grade km rats with two different parameters of electroporation (low voltage 30 ~ 110V / cm) and high voltage electric perforation (500V / cm) to observe the effects of different electroporation conditions on the degree of testicular injury and reproductive ability of SPF km male rats. In order to screen the appropriate parameters of electroporation in vivo. On the basis of this, we treated the eukaryotic expression plasmid pCE-29DNA with enhanced green fluorescent protein (EGFP) gene by two different treatments: 1) pCE-29DNA transfection reagent in vivo jetPII (TMTM) Trypan blue TBX 2) pCE-29DNA Trypan blue TBN. And then put the two different
【學(xué)位授予單位】：第一軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R-332

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 沈新明,喬貴林,張玲,江培洲,黃華,姚開泰;用曲細(xì)精管微注射法建立綠色熒光蛋白轉(zhuǎn)基因小鼠[J];第一軍醫(yī)大學(xué)學(xué)報(bào);2002年03期

2 張繼峰,秦陽(yáng)君,付愛(ài)華,湯健,陳光慧,蔡?hào)|,韓濟(jì)生;電針介導(dǎo)的基因轉(zhuǎn)移[J];中國(guó)科學(xué)C輯;1999年01期

3 郭秀洋,周澤揚(yáng),馮麗春,汪琳,魯成,向仲懷;利用精子介導(dǎo)法向蠶卵導(dǎo)入外源基因的研究[J];生物化學(xué)與生物物理進(jìn)展;2001年03期

4 楊凱,程漢華,郭一清,周榮家;采用高分子介導(dǎo)精子作載體制備轉(zhuǎn)基因泥鰍[J];遺傳學(xué)報(bào);2001年12期

，

本文編號(hào)：1944022