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DIFFERENTIAL EXPRESSION MECHANISMS OF HOUSEKEEPING AND LUXURY GENES DURING EMBRYONIC AND POST-EMBRYONIC DEVELOPMENT

管家基因和奢侈基因在胚胎及胚胎后发育过程中的差异表达机制

基本信息

批准号：
08406006
负责人：
KOGA Katsumi
金额：
$ 24.38万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1999
项目状态：
已结题

项目摘要

The gene expression processes in Bombyx mori were examined in terms of genetics, morphology, biochemistry and molecular biology and results obtained were comprehensively debated. First, the organ-specific expression of aldolase isozymes, as a typical housekeeping gene system, was analyzed. Two isozymes we have found were purified and the mode of occurrence of mRNA during embryogenesis was demonstrated using a cDNA clone obtained, confirming the switchover of aldolase isozymes at a critical period of embryogenesis shortly before hatching. Moreover, the enzymatic properties of the silkworm aldolases were compared with those from the housefly, human and lamprey also investigated in the present study. Second, the luxurious multigene families for the humoral trypsin inhibitors of B. mori were successfully analyzed. In particular of interest were our pursuit for the gene network underlying the spermatogenesis and homologous recombination ; the subtraction method of cDNAs for the proteins interacting with DMC1 led us to isolate clones encoding cytoplasmic and mitochondrial ATPase inhibitor proteins, G-protein regulatory protein etc., for the first time in higher eukaryotes. Third, the B. mori mutant lines in particular those related to embryogenesis, stored in Kyushu University, were investigated for morphology and embryonic protein constitutions. We also analyzed the chromosome number dimorphism using many lines of Bombyx mandarina, which is putative ancestor of B. mori, and confirmed the geographical borderline. As a whole the results obtained in the present project have verified the status of B. mori as a modern model system of molecular and evolutional biology.

从遗传学、形态学、生物化学和分子生物学等方面对家蚕基因表达过程进行了研究，并对所获得的结果进行了综合讨论。首先，对典型的看家基因系统--醛缩酶同工酶的器官特异性表达进行了分析。对我们发现的两种同工酶进行了纯化，并用获得的一个克隆证实了胚胎发生过程中信使核糖核酸的存在方式，证实了在胚胎发生的关键期孵化前的醛缩酶同工酶发生了转换。此外，还与家蝇、人和七鳃鳗的醛缩酶进行了比较。其次，成功地分析了家蚕体液胰蛋白酶抑制剂的奢侈多基因家族。尤其令人感兴趣的是我们对精子发生和同源重组背后的基因网络的追求；与DMC1相互作用的蛋白质的cDNA的消减方法使我们首次在高等真核生物中分离到编码细胞质和线粒体ATPase抑制蛋白、G蛋白调节蛋白等的克隆。第三，对保存在九州大学的家蚕突变系，特别是与胚胎发生相关的突变系进行了形态和胚胎蛋白质组分的研究。此外，我们还对野桑蚕的多个品系进行了染色体数目二态分析，确定了野桑蚕的地理分界线。作为一个整体，本项目所获得的结果验证了家蚕作为分子和进化生物学的现代模式系统的地位。