水稻Rubisco小亚基前体cDNA的克隆及其产物向豌豆叶绿体的运输  

Cloning of Rice rbcs Precursor cDNA and the Import of Its in vitro Synthesised Products into Intact Chloropasts from Pea

作　　者：陈根云[1] 颜日辉 李立人[1] 

机构地区：[1]中国科学院上海植物生理研究所,上海200032

出　　处：《植物生理学报（0257-4829)》1998年第3期293-299,共7页Acta Phytophysiologica Sinica

基　　金：国家自然科学基金;中国科学院"九五"基础性研究重点项目

摘　　要：用RT-PCR方法克隆了完整的水稻Rubisco小亚基前体cDNA基因，经耦联的体外转录和翻译系统合成了带同位素标记的小亚基前体蛋白，然后与新制备的豌豆完整叶绿体共保温，进行蛋白质的跨膜运输研究显示：异源的水稻Rubisco小亚基前体能穿膜运输入豌豆叶绿体。成熟小亚基不能进入叶绿体，进入叶绿体的小亚基量在一定范围内与外加的小亚基前体量成正比，光对小亚基的跨膜运输有一定的促进作用，外加ATP能显著促进小亚基前体的运输，而ADP无此作用。The Precurso cDNA of Rubisco small subunit (rbcS) from rice was cloned in pSPT19 by RT-PCR The results of sequencing indicated that the fragment of rbcS contained the full length transit peptide. By comparing its sequence of transit peptide with that of Pea, it was foun that the Percentage of homolgy of the transit peptide sequence between the two species is only 39. 5 %, but they have three high conservation re gions, in which the percentag of homologues is up to 80%. Furthermore, the rice precursor of rbcS (PrSS) labeled with 35S-Met was Synthsised in vitro from plasmid-transcribed mRNA in wheat germ system(cell free). Afte being incubated with hesh intact chloroplasts from pea, rice prSS was shown to be able to cross the mermbran of Pea chloroplasts. the amoun of the PrSS imported into choloroplast under illumination was 17% more than that treated in the dark; and the amount was increased by 80% when the chlorplasts is treasted withATP, ADP had no obvious effect on the import Furthermore, the amoun of SS imported into the chloroplasts was directly proportional to the amount of PrSS added in the import assays.

关 键 词：RUBISCO小亚基 RT-PCR 体外转录/翻译 叶绿体 

分 类 号：Q732[生物学—分子生物学] Q945