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研究生:程偉政
論文名稱:早期小鼠胚表現之新同源箱基因Obox6分子特性探討
論文名稱(外文):Molecular characterization of Obox6, a novel homeobox gene preferentially expressed in early embryonic stages
指導教授:李鴻李鴻引用關係沈哲鯤沈哲鯤引用關係
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:遺傳學研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:2003
畢業學年度:91
語文別:中文
中文關鍵詞:同源箱基因同源功能區小鼠胚發育Obox同源箱基因家族二細胞胚桑椹胚基因剔除小鼠
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同源箱基因在生物的發育過程中扮演一個重要的角色,其胺基酸序列在演化上具有高度的保守性,本試驗中,即針對同源功能區部分設計了退化性引子,於小鼠胚二細胞時期的cDNA library篩選出新的同源箱基因─Obox6基因(oocyte specific homeobox gene),經電腦分析比對之後,得知Obox6基因座落在小鼠第七對染色體上,橫跨約1.5 kb的區域,包含兩個表現子(exon)及一個介入子(intron),其同源箱序列分割在兩個表現子內。Obox6基因可轉譯出由347個胺基酸組成的蛋白質,其中同源功能區之胺基酸序列和Obox同源箱基因家族者具高度的相似性。
利用RT-PCR分析Obox6基因在mRNA層次的表現,則可發現其在小鼠早期胚的發育時期有專一性的表現,特別是在小鼠胚發育至二細胞胚與桑椹胚期之間表現量更為顯著。將His6-Obox6融合蛋白質用以製備Obox6多株抗體,此多株抗體對Obox6蛋白質具有高度的專一性,在免疫細胞化學分析法的試驗結果中再次證明Obox6蛋白質確實存在於早期小鼠胚中,其在發育至囊胚期時則呈明顯減少的現象。
綜合以上結果推測Obox6基因可能對於小鼠早期胚發育具有重要的影響。為了進一步研究此基因的功能,首先採用基因剔除技術,產製Obox6基因剔除小鼠,目前已成功得到三株Obox6基因剔除的胚幹細胞株,並利用囊胚顯微注射技術得到八隻基因嵌合鼠,未來將可藉由分析Obox6基因剔除小鼠的表現型,進而提供此基因對於影響小鼠早期胚胎發育的重要證據。
目錄
英文摘要………………………………………………………………1
中文摘要………………………………………………………………2
Ⅰ. 序論………………………………………………………………3
Ⅱ. 實驗材料與方法 …………………………………………………8
Ⅲ. 實驗結果…………………………………………………………18
Ⅳ. 討論………………………………………………………………22
Ⅴ. 圖…………………………………………………………………26
Ⅵ. 附錄………………………………………………………………39
Ⅶ. 參考文獻…………………………………………………………47
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