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

參考文獻
Almouzni, G. and A.P. Wolffe. 1995. Constraints on transcriptional activator function contribute to transcriptional quiescence during early Xenopus embryogenesis. EMBO J. 14: 1752-1765
Bachvarova, R.F. 1992. A maternal tail of poly(A): the long and the short of it. Cell 69: 895-897.
Bellier, S., S. Chastant, P. Adenot, M. Vincent, J.P. Renard, and O. Bensaude. 1997. Nuclear translocation and carboxyl-terminal domain phosphorylation of RNA polymerase II delineate the two phases of zygotic gene activation in mammalian embryos. EMBO J. 16: 6250-6262.
Bouniol, C., E. Nguyen, and P. Debey. 1995. Endogenous transcription occurs at the 1-cell stage in the mouse embryo. Exp. Cell Res. 218: 57-62.
Bürglin, T.R. 1997. Analysis of TALE superclass homeobox genes (MEIS, PBC, KNOX, Iroquois, TGIF) reveals a novel domain conserved between plants and animals. Nucleic Acids Research. 25: 4173-4180.
Doe, C.Q., Y. Hiromi, W.J. Gehring, and C.S. Goodman. 1988. Expression and function of the segmentation gene fushi tarazu during Drosophila neurogenesis. Science 239: 170-175.
Duboule, D. and G. Morata. 1994. Colinearity and functional hierarchy among genes of the homeotic complexes. Trends Genet. 10: 358-364.
Duboule, D. 1994. Guidebook to the Homeobox Genes. First editiion.
Dyce, J., M. George, H. Goodall ,and T.P. Fleming. 1987. Do trophectoderm and inner cell mass cells in the mouse blastocyst maintain discrete lineages? Development 100: 685-698.
Gehring, W.J. 1987. Homeo boxes in the study of development. Science 236: 1245-1252.
Gilbert, S.F. 2000. Development biology, Sixth edition.
Hogen, B., F. Costantini, E. Lacy, and R. Beddington. 1994. Manipulating the Mouse Embryo：a laboratory manual, Second edition.
Kanka, J. 2003. Gene expression and chromatin structure in the pre-implantation embryo. Theriogenology 59: 3-19.
Laughon, A. and M.P. Scott. 1984. Sequence of a Drosophila segmentation gene: protein structure homology with DNA-binding proteins. Nature 310: 25-31.
McGinnis,W., M.S. Levine, E. Haefen, A. Kuroiwa, and W.J. Gehring. 1984a. A conserved DNA sequence in homeotic genes of the Drosophila Antennapedia and bithorax complexes. Nature 308: 428-433.
McGinnis,W., R.L. Garber, J. Wirz, A. Kuroiwa, and W.J. Gehring. 1984b. A homologous protein-coding sequence in Drosophila homeotic genes and its conservation in other metazoans. Cell 37: 403-408.
Mortensen, R.M., M. Zubiaur, E.J. Neer, and J.G. Seidman. 1991. Embryonic stem cells lacking a functional inhibitory G-protein subunit (αi2) produced by gene targeting of both alleles. Proc. Natl. Acad. Sci. U.S.A. 88: 7036-7040.
Nothias, J.Y., S. Majunder, K.J. Kaneko, and M.L. DePamphilis. 1995. Regulation of gene expression at the beginning of mammalian development. J. Biol. Chem. 270(38): 22077-22080.
Otting, G., Y.Q. Qian, M. Billeter, M. Muller, M. Affolter, W.J. Gehring, and K. Wuthrich. 1990. Protein-DNA contacts in the structure of a homeodomain-DNA complex determined by nuclear magnetic resonance spectroscopy in solution. EMBO J. 9: 3085-3092.
Paynton, B.V., R. Rempel, and R. Bachvarova. 1988. Changes in state of adenylation and time course of degradation of maternal mRNAs during oocyte maturation and early embryonic development in the mouse. Dev. Biol. 89: 363-378.
Pedersen, R.A., K. Wu, and H. Batakier. 1986. Origin of the inner cell mass in the mouse embryos: Cell lineage analysis by microinjection. Dev. Biol. 117: 581-595.
Percival-Smith, A., M. Muller, M. Affolter, and W.J. Gehring. 1990. The interaction with DNA of wild-type and mutant fushi tarazu homeodomains. EMBO J. 9: 3967-3974.
Prioleau, M.N., J. Huet, A. Sentenac, and M. Mechali. 1994. Competition between chromatin and transcription complex assembly regulates gene expression during early development. Cell 77: 439-449.
Rajkovic, A., C. Yan, W. Yan, M. Klysik, and M.M. Matzuk. 2002. Obox, a family of homeobox genes preferentially expressed in germ cells. Genomics 79(5): 711-717.
Schultz, G.A. and S. Heyner. 1992. Gene expression in pre-implantation mammalian embryos. Mutat. Res. 296: 17-31.
Schultz, R.M. 1993. Regulation of zygotic gene activation in the mouse. Bioessays 1993: 531-538.
Schultz, R.M., W.Jr. Davis, P. Stein, and P. Svoboda. 1999. Reprogramming of gene expression during preimplantation development. J. Exp. Zool. 285: 276-282.
Stanton, J.A., A.B. Macgregor, and D.P. Green. 2003. Gene expression in the mouse preimplantation embryo. Reproduction 125: 457-468.
Wassarman, P.M., and R.A. Kinloch. 1992. Gene expression during oogenesis in mice. Mutat. Res. 296: 3-15.
Wolpert, L. 1998. Principles of development. First edition.
Worrad, D.M., P.T. Ram, and R.M. Schultz. 1994. Regulation of gene expression in the mouse oocyte and early implantation embryo: developmental changes in Sp1 and TATA box-binding protein, TBP. Development. 120: 2347-2357.