Schrader S, Johanningmeier U
Lehrstuhl für Biochemie der Pflanzen, Ruhr-Universität Bochum, Germany.
Plant Mol Biol. 1992 May;19(2):251-6. doi: 10.1007/BF00027346.
The D1-precursor protein of the photosystem II reaction centre contains a carboxy-terminal extension whose proteolytic removal is necessary for oxygen-evolving activity. To address the question of the role of the carboxy-terminal extension in the green alga Chlamydomonas reinhardtii, we truncated D1 by converting codon Ser345 of the psbA gene into a stop codon. Particle gun transformation of an in vitro modified psbA gene fragment also carrying mutations conferring herbicide resistance yielded a homoplasmic transformant containing the stop codon. Since oxygen evolution capacity is not affected in this mutant as compared with herbicide-resistant control cells, the carboxy-terminal extension is dispensable for a functional photosystem II complex under normal growth conditions.
光系统II反应中心的D1前体蛋白含有一个羧基末端延伸序列,其蛋白水解去除对于放氧活性是必需的。为了解决羧基末端延伸序列在莱茵衣藻中的作用问题,我们通过将psbA基因的密码子Ser345转换为终止密码子来截短D1。对还携带赋予除草剂抗性突变的体外修饰psbA基因片段进行粒子枪转化,得到了含有终止密码子的同质转化体。由于与抗除草剂对照细胞相比,该突变体的放氧能力不受影响,因此在正常生长条件下,羧基末端延伸序列对于功能性光系统II复合体是可有可无的。