野生型衣藻中光系统II D1亚基的位点特异性诱变。
Site-specific mutagenesis of the D1 subunit of photosystem II in wild-type Chlamydomonas.
作者信息
Przibilla E, Heiss S, Johanningmeier U, Trebst A
机构信息
Ruhr-Universität Bochum, Lehrstuhl für Biochemie der Pflanzen, Federal Republic of Germany.
出版信息
Plant Cell. 1991 Feb;3(2):169-74. doi: 10.1105/tpc.3.2.169.
Abstract
The structure and functional mode of photosystem II reaction center protein D1 can be studied by analyzing the effects of amino acid substitutions within the binding niche for QB, the second stable electron acceptor of photosystem II, on herbicide binding. Here we report on site-directed mutagenesis of the psbA gene coding for the D1 protein in the unicellular alga Chlamydomonas reinhardtii. The chloroplasts of wild-type cells were transformed using the particle gun. The plasmids introduced carried an in vitro mutated fragment of the psbA gene. We obtained a double mutant with replacements of amino acids 264 and 266 and a triple mutant having an additional substitution in position 259. The sensitivities of both mutants toward several types of herbicides are given and compared with those of a mutant having only a substitution at position 264.
摘要
通过分析光系统II（PSII）的第二个稳定电子受体QB结合位点内氨基酸取代对除草剂结合的影响，可以研究光系统II反应中心蛋白D1的结构和功能模式。在此，我们报道了对单细胞绿藻莱茵衣藻中编码D1蛋白的psbA基因进行定点诱变的研究。使用粒子枪对野生型细胞的叶绿体进行转化。导入的质粒携带psbA基因的体外突变片段。我们获得了一个氨基酸264和266被替换的双突变体以及一个在位置259处有额外取代的三突变体。给出了这两个突变体对几种除草剂的敏感性，并与仅在位置264处有取代的突变体进行了比较。
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