Cloning, expression of the psbU gene, and functional studies of the recombinant 12-kDa protein of photosystem II from a red alga Cyanidium caldarium.
Cloning, expression of the psbU gene, and functional studies of the recombinant 12-kDa protein of photosystem II from a red alga Cyanidium caldarium.
复制标题
psbU 基因的克隆、表达以及红藻 Cyanidium caldarium 光系统 II 重组 12-kDa 蛋白的功能研究。
DOI：
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发表时间：
1999
期刊：
影响因子：
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通讯作者：
I. Enami
中科院分区：
文献类型：
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作者：
H. Ohta;A. Okumura;Satoshi Okuyama;Ai Akiyama;Masako Iwai;Shizue Yoshihara;Jian;M. Kamo;I. Enami
利用PCR和RACE技术克隆了一种红藻（Cyanidium caldarium）PS Ⅱ复合物的外源12 kDa编码蛋白，并进行了序列测定。该基因编码一个154个氨基酸的多肽，计算分子量为16，714 Da。该蛋白的全序列包括两个特征性转运肽，一个用于跨叶绿体被膜转运，另一个用于靶向进入类囊体腔。这表明该蛋白质在核基因组中编码。该成熟蛋白由93个氨基酸组成，分子量为10，513 Da，在大肠杆菌中成功表达，纯化后与33和20 kDa的外源蛋白及cyt c-550重组为CaCl 2洗涤的PS II复合物。重组的12-kDa蛋白完全与PSII复合物结合，这导致氧释放的恢复等于通过结合天然12-kDa蛋白所达到的水平。
The encoding extrinsic 12-kDa protein of oxygen-evolving PS II complex from a red alga, Cyanidium caldarium, was cloned and sequenced by means of PCR and a rapid amplification of cDNA ends (RACE) procedure. The gene encodes a putative polypeptide of 154 amino acids with a calculated molecular mass of 16,714 Da. The full sequence of the protein includes two characteristic transit peptides, one for transfer across the chloroplast envelope and another for targeting into the thylakoid lumen. This indicates that the protein is encoded in the nuclear genome. The mature protein consists of 93 amino acids with a calculated molecular mass of 10,513 Da. The cloned gene was successfully expressed in Escherichia coli and the resulting protein was purified, reconstituted to CaCl2-washed PS II complex together with the other extrinsic proteins of 33 and 20 kDa and cyt c-550. The recombinant 12-kDa protein bound completely with the PSII complex, which resulted in a restoration of oxygen evolution equal to the level achieved by binding of the native 12-kDa protein.
影响因子：
11.4
作者：
LUTCKE, HA;CHOW, KC;SCHEELE, GA
通讯作者：
SCHEELE, GA