Molecular and genetical research developmental processes in ciliates

1. 通过对其发展过程的分析，得出了主要结论。(1)在偶联过程中，发现Mic分裂诱导因子（Mic DIF）和Mic DIF抑制因子在非常特定的阶段表达。在减数分裂早期进行热处理可引起这些因素的抑制。(2)发现抗巨核单克隆抗体可识别核仁蛋白。分离到识别减数分裂微核、可能的配对型物质V和位于口腔侧的纤毛的单克隆抗体。(3)恢复jumyo突变体正常生长速率的草履虫生长因子（ParGF）在功能上与人胎血清相似。(4)原认为与配子体核形成有关的14nm细丝被鉴定为线粒体柠檬酸合成酶。分离到一个与14nm长丝结合的新蛋白，鉴定为延伸因子-1alpha。(5)不成熟期短的变异昼夜节律短。生理节律与生理活动之间存在相关性。成熟细胞注射了未成熟细胞的DNA后变得不成熟。这表明发育时钟本身在发育过程中涉及DNA的变化。(6)共种间大核浆移植发现了配型控制基因的同种特异性转化。突变基因转化或克隆的发展。(1)将野生型消化的DNA片段导入突变细胞后，10kb以上的DraI片段能有效恢复tnd2突变体。如果能找到合适的酶，就有可能克隆出tnd2基因。(2)从stylonychia衍生的载体Tubingen3可用于四膜虫和草履虫。(3)草履虫线粒体中存在非细菌包涵体和质粒样DNA。通过DNA分析，质粒样DNA分为三组。

1. Main results obtained from the analyzes of the developmental processes.(1) Mic division inducing facor (mic DIF) and suppressor for the mic DIF were found expressed at very distict stage during conjugation process, . Heat treatment at early stage of meiosis induced an arrest caused by the factors. (2) Monoclonal antibodies against a macronucleus were found to recognize a nucleolar protein. Monoclonal antibodies which recognize the micronucleus during meiosis, possible mating type substance V and cilia located at oral side were isolated. (3) ParGF(Paramecium grouth factor) which rescues normal growth rate of jumyo mutant exhibited functional similarity to human fetal serum. (4) The 14nm filament which has been though to be involving in the formation of gametic nuclei, was identified as the mitochondrial citrate synthetase. A new protein which copurifies with the 14nm filament was isolated and identified as the elongation factor-1alpha. (5) A variant with short immaturity period had a short circadian rhythm. A corelation between circadian rhythm and phisiologica activity was observed. Mature cells injected with DNA from immature cells become immature. This suggests that a developmental clock itself involves a change of DNA during development. (6) Conversions of syngen specificity of the mating type controlling genes were discovered by intersyngenic macronucleoplasmic transplantation.2. Development of transformation or cloning of mutated genes. (1) DraI fragments more than 10kb were effective to restore the tnd2 mutants when digested DNA fragments from wild type were introduced to mutant cells. If the proper enzyme will be found, cloning of tnd2 gene may be possible. (2) Tubingen3, the vector derived from stylonychia can be used for both Tetrahymena and Paramecium. (3) Non-bacterial inclusion bodies and plasmid-like DNA in mitochondria were found in Paramecium. By DNA analyzes, the plasmid-like DNAs were deviding to three groups.