FUNCTION OF THE KINETOPLAST IN THE HEMOFLAGELLATES

这个项目的总体目标是剖析线粒体的过程。 发生在低等真核寄生细胞中的分子生物发生 就线粒体基因组和核基因组而言。 此外，我们还想了解这种不寻常的分子 线粒体DNA在结构和功能上的分化 有道理。具体目标如下：1.完成全序列 马尾藻L.tarentolae Maxicircle.2.所有3‘及5’端的识别 成绩单。3.初级成绩单的鉴定。4.与世隔绝 对tRNA进行测序。5.特定上颌骨基因的转录 不同的生理条件。6.上颌骨基因的转录 在布氏松毛虫生活史上。7.线粒体RNA聚合酶的分离。 上颌骨基因启动子的鉴定。8.比较上颌骨 不同物种的基因组织。9.9和12S基因的测序 来自几个物种。10.细胞色素氧化酶或FOF1 ATPase的分离 从狼毒乳杆菌中分离得到。11.细胞色素氧化酶核基因的克隆 FOF1 ATPase亚基的4-7亚基。12.克隆核基因 线粒体蛋白质。13.对发育调节的分析 布氏毛滴虫Bhr序列的转录。14.对可能性的分析 微环转录。

The overall goal of this project is to dissect the process of mitochondrial biogenesis that occurs in this lower eukaryotic parasitic cell in molecular terms, both in terms of the mitochondrial genome and the nuclear genome. In addition we would like to understand the unusual molecular differentiation of this mitochondrial DNA in a structural and functional sense. Specific aims are as follows: 1. Completion of entire sequence of L. tarentolae maxicircle. 2. Identification of 3' and 5' ends of all transcripts. 3. Identification of primary transcripts. 4. Isolation and sequencing of tRNAs. 5. Transcription of specific maxicircle genes under different physiological conditions. 6. Transcription of maxicircle genes in life cycle of T. brucei. 7. Isolation of mitochondrial RNA polymerase. Identification of promoters of maxicircle genes. 8. Comparative maxicircle gene organization in different species. 9. Sequencing of 9 and 12s genes from several species. 10. Isolation of cytochrome oxidase or FOF1 ATPase from L. tarentolae. 11. Cloning of nuclear genes for cytochrome oxidase subunits 4-7 of FOF1 ATPase subunits. 12. Cloning nuclear genes for mitochondrial proteins. 13. Analysis of developmentally regulated transcription of the BhR sequence in T. brucei. 14. Analysis of possible minicircle transcription.