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铜绿微囊藻生物钟蛋白KaiC的自激活活性和自身相互作用研究 被引量:3
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作者 王靖 徐虹 +2 位作者 李珣 郑锦乾 王立红 《海洋环境科学》 CAS CSCD 北大核心 2008年第1期67-70,82,共5页
通过PCR扩增了铜绿微囊藻的生物钟主控基因KaiC,并将其分别克隆到酵母双杂交系统的诱饵质粒pGBKT7和猎物质粒pGADT7中,然后将重组质粒pGBKT7-kaiC/pGADT7-kaiC和pGBKT7-kaiC/pGADT7分别共转化酵母菌AH109,经营养缺陷生长和β-半乳糖苷... 通过PCR扩增了铜绿微囊藻的生物钟主控基因KaiC,并将其分别克隆到酵母双杂交系统的诱饵质粒pGBKT7和猎物质粒pGADT7中,然后将重组质粒pGBKT7-kaiC/pGADT7-kaiC和pGBKT7-kaiC/pGADT7分别共转化酵母菌AH109,经营养缺陷生长和β-半乳糖苷酶印迹检测表明,KaiC蛋白不具有毒性不会影响酵母细胞的生长,也不具有自激活活性,不会激活报告基因的表达,并且KaiC蛋白自身存在较强的相互作用。诱饵质粒pGBKT7-kaiC可用于从基因组文库中筛选KaiC的相互作用蛋白。 展开更多
关键词 铜绿微囊藻 钟蛋白kaic 酵母双杂交
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Isolation, Purification and Characterization of Nucleoids from <i>Synechococcus elongatus</i>PCC 7942
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作者 Ali Azam Talukder Takao Kondo 《Advances in Microbiology》 2014年第15期1105-1116,共12页
The genomic DNA of bacteria is highly compacted in one or a few bodies known as nucleoids. In order to understand the overall configuration and physiological activities of the cyanobacterial nucleoid under various gro... The genomic DNA of bacteria is highly compacted in one or a few bodies known as nucleoids. In order to understand the overall configuration and physiological activities of the cyanobacterial nucleoid under various growth conditions and the role(s) of each nucleoid protein in clock function, thylakoid membrane-associated nucleoids from the Synechococcus elongatus (se) PCC 7942 strain were isolated and purified in presence of spermidine at low salt concentrations by sucrose density gradient centrifugation. The sedimentation rates, protein/DNA composition and microscopic appearances as well as variation in structural components of clock proteins from the isolated nucleoids were compared under identical conditions. Microscopic appearances of the nucleoids were consistent with the sedimentation profiles. The nucleoid structure in the wild type was more tightly compacted than that in the KaiABC mutant strain. Western immunoblot analyses revealed that the KaiC was associated with the nucleoid fraction whereas maximum KaiA was localized in the cytosolic fraction, supposedly in association with the translation machinery. 展开更多
关键词 SYNECHOCOCCUS elongatus Sucrose Gradient CYANOBACTERIAL NUCLEOIDS Circadian Clock kaic protein
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生物钟翻译后核心振荡机制 被引量:3
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作者 杨丽婷 刘森 《生命的化学》 CAS CSCD 2016年第3期390-396,共7页
生物钟现象是普遍存在于生物体内的一种周期约24小时的节律性振荡现象。就生物钟的振荡机制而言,依赖于转录-翻译的负反馈回路(transcriptional/translational feedback loop,TTFL)机制,这也是最经典的振荡机制。但是近些年来,众多新的... 生物钟现象是普遍存在于生物体内的一种周期约24小时的节律性振荡现象。就生物钟的振荡机制而言,依赖于转录-翻译的负反馈回路(transcriptional/translational feedback loop,TTFL)机制,这也是最经典的振荡机制。但是近些年来,众多新的研究发现,生物钟翻译后振荡(post-translational oscillation,PTO)机制也具有重要作用,并在古生菌、原核生物和真核生物中都有发现。目前研究最深入的生物钟PTO是原核生物蓝藻中由Kai A/Kai B/Kai C三个蛋白构成的生物钟振荡。近几年来,一类普遍存在于几乎所有生物体内的过氧化物氧还蛋白(peroxiredoxin,PRX)依赖的生物钟振荡,也取得了一系列的重要研究进展。本文对近些年来生物钟PTO的研究进行了综述,并介绍了PTO与TTFL两种生物钟机制的相互联系。 展开更多
关键词 转录—翻译负反馈回路 翻译后振荡 kaic 过氧化物氧还蛋白 蛋白质相互作用
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