Molecular manipulation and application of thermophilic cyanobacteria

建立了嗜热蓝藻细长嗜热菌BP-1的转化能力克隆。我们与Tabata(Kazusa DNA Res.Inst.)合作，测定了该基因的全序列，克隆了一个新的黄素结合蛋白基因，并在大肠杆菌中进行了表达，分离得到了藻蓝蛋白，并进行了结晶。获得了2.0A分辨率的X射线衍射谱数据，并对长穗拟青霉中的pSBI、psbT、psbV和psbK基因进行了突变。从这些突变体中分离出光系统II复合体，构建了一系列针对长穗金龟子外源33 kDa蛋白的定点突变体，并对其进行了功能分析；从野生株中分离出放氧光系统II复合体并结晶。3D

A transformation-competent clone of the thermophilic cyanobacterium Thermosynechococcus elongatus strain BP-1 was established. In collaboration with S. Tabata (Kazusa DNA Res. Inst.), we determined the complete nucleotide sequence.A gene for a novel flavin-binding protein was cloned from T. elongatus and expressed in E. coli.Phycocyanin was isolated from T. elongatus and crystalled. X-ray diffraction data at 2.0 A resolution was obtained.Gene disruption mutants were created for psbI, psbT, psbV and psbK in T. elongatus. Photosystem II complexes were isolated from these mutants.A number of site-directed mutants were constructed for the extrinsic 33 kDa protein of T. elongatus and were analyzed functionally.O_2-evolving Photosystem II complex was isolated from wild type of T. vulcanus and crystallized. The 3D