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编码Synechococcus sp. PCC 7002 FNR 中FNR区的基因克隆及其在大肠杆菌中的表达 被引量:3

Cloning of the Gene Encoding FNR Domain of FNR of Cyanobacterium Synechococcus sp. PCC 7002 and its Overexpression in Escherichia coli 
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摘要 用PCR的方法克隆出了编码蓝细菌Synechococcussp.PCC7002FNR中FNR区的基因petHL,克隆到达载体pET3a上,转化大肠杆菌BL21(DE3)后实现了大量表达。重组FNR区(rFNRD)经DEAESephdexA50离子交换层析及SephadexG100凝胶层析得到大量的电泳均一的rFNRD。N末端氨基酸序列分析表明,表达产物确为petHL所编码。且起始Met翻译后未被除去。rFNRD与rFNR的吸收光谱相同,其黄递酶活性的最适pH和最适温度也相同。 The petHL of Synechnococcus sp. PCC 7002 encoding FNR domain (FNRD) of FNR was amplified by PCR, and cloned into expressing vector pET 3a. Overexpression of petHL was achieved with E. coli BL21 (DE3). The recombinant FNRD was purified to homogeneity by DEAE Sephadex A 50 and Sephadex G 100 chromatography. N terminal amino acid sequencing showed that rFNRD was encoded by petHL and initial Met was not posttranslationally removed. rFNRD had the same absorption spectrum, optimal pH and optimal temperature as those of rFNR. rFNRD could catalyze photosynthetic electron transport from P700 to NADP + in vitro.
出处 《Acta Botanica Sinica》 CSCD 1999年第4期389-392,共4页 Acta Botanica Sinica(植物学报:英文版)
基金 国家自然科学基金
关键词 蓝细菌 FNR区 高效表达 电子传递 PCR Cyanobacteria FNR domain Overexpression Electron transport
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参考文献3

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同被引文献19

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