發(fā)布時間：2018-05-27 22:19

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

【摘要】：轉基因動物在醫(yī)學、生命科學、生物制藥等領域有著廣泛的研究和應用前景,因此科學家們一直在探索一種更簡便易行、經(jīng)濟實用而又高效的轉基因方法。近幾年來,隨著科學技術的發(fā)展,體內基因轉移已成為用于基因治療、生物學分析等領域非常受歡迎的一種技術,體內電穿孔技術就是其中之一。 精原干細胞是精子的前體細胞,是雄性動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胞類型。對精原干細胞的操作和修飾在建立轉基因動物模型、制備乳腺生物反應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并進行體內電穿孔以建立轉基因動物的可行性,我們首先設計了低壓(30～110V/cm)和高壓(500～900V/cm)兩組不同的體內電穿孔參數(shù)對SPF級KM雄鼠睪丸進行在體電穿孔,以觀察不同的電穿孔條件對SPF級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電穿孔參數(shù)。并在此基礎上,我們對插入有增強型綠色熒光蛋白(EGFP)基因的真核表達質粒pCE-29DNA進行2種不同的處理:(1) pCE-29DNA+轉染試劑(in vivo jetPI~(TM)+臺盼藍(TB);(2) pCE-29DNA+臺盼藍(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
【學位授予單位】：第一軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2005
【分類號】：R-332

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本文編號：1944022