转小鼠金属硫蛋白基因集胞藻6803及其野生藻培养

通过摇瓶培养考察转小鼠金属硫蛋白基因集胞藻６８０３及其野生藻的光合自养培养过程中碳源浓度、氮源浓度和光照度对这两种藻生长的影响。野生藻的最适生长碳源浓度（０．０５－０．５ｇ／Ｌ）比转基因藻的最适生长碳源浓度（０．０２－０．０５ｇ／Ｌ）高。氮源浓度对转基因藻和野生藻的影响趋势相同。在０．３－４．５ｇ／Ｌ范围内,随着氮源浓度的增高,藻体生长的最大细胞浓度降低。转基因藻生长的最佳光照度为１５００ｌｘ,野     

Sodium Acetate Stimulates PHB Biosynthesis in Synechocystis sp. PCC 6803

IntroductionA large variety of bacteria are able to synthesizepolyhydroxyalkanoic acids (PHAs) and to depositlarge amounts of these polyesters as insolubleinclusions in cytoplasm[1 ] . PHAs have attractedthe interest of the chemical industry because oftheir biodegradable properties for applications invarious technical,medical,and pharmaceuticalapplications[2 ] .Poly-β-hydroxybutyrate(PHB) isatypical PHA that is produced by heterotrophicbacteria,such as Alcaligenes eutrophus andEscherichi…     

Two-Component Signal Transduction Systems in the Cyanobacterium Synechocystis sp. PCC 6803

Introduction Rapid adaptation to environmental challenges is vitally important for bacterial survival and growth. One way in which bacteria control their response to changing environmental conditions is through the mechanism of two-component signal transd…     

集胞藻PCC6803基因ssl1690缺失突变株的构建

使用BLAST软件对聚球藻PCC7002的裂合酶基因cpcT进行了同源搜索分析,在集胞藻PCC6803中获取了相似程度达68%的同源基因ssl1690.为进一步探讨ssl1690功能,构建了同源缺失突变载体,将其转入蓝藻细胞中,筛选得到基因缺失突变株,并将基因缺失突变株培养于BG11液体培养基中,观察了其生长情况和表型变化.结果表明:基因缺失突变株与野生株相比,生长有所延迟,有漂白现象.通过分离和比较SDS-PAGE电泳突变株与野生株的藻胆体蛋白,发现缺失株存在藻胆体组分蛋白的缺失.故认为集胞藻PCC6803 ssl1690基因功能与光合作用有关,其所编码的蛋白对藻蓝蛋白正常的体内生物合成具有重要作用.     

Bifunctional enzyme FBPase/SBPase is essential for photoautotrophic growth in cyanobacterium Synechocystis sp. PCC 6803

From a random insertion mutant library of Synechocystis sp. PCC 6803, a mutant defective in photoautotrophic growth was obtained. The interrupted gene was identified to be slr2094 （fbpl）, which encodes the fructose-1,6-biphosphatase （FBPase）/sedoheptulose-1,7-biphosphatase （SBPase） bifunctional enzyme （F-I）. Two other independently constructed slr2094 mutants showed an identical phenotype. The FBPase activity was found to be virtually lacking in an slr2094 mutant, which was sensitive to light under mixotrophic growth conditions. These results indicate that slr2094 is the only active FBPase-encoding gene in this cyanobacterium. Inactivation of photosystem II by interrupting psbB in slr2094 mutant alleviated the sensitiveness to light. This report provides the direct genetic evidence for the essential role of F-I in the photosynthesis of Synechocystis sp. PCC 6803.     

Contributions of DPOR at Low Light Intensi.ty to Chlorophyll Biosynthesis and Growth in the Synechocystis sp. PCC 6803

Introduction Chlorophylls (Chls) and bacteriochlorophylls (Bchls) are two of the most abundant classes of natural pigments, and their biosynthesis is a major metabolic activity in the ecosystem[1]. Protochlorophyllide (Pchlide) is an intermediate in the Chl and Bchl biosynthetic pathway of photosynthetic organisms and its reduction to chlorophyllide (Chlide) is a key regulatory step in the Chl biosynthesis pathway. In this reaction, the porphyrin D-ring of Pchlide is converted to chlorin b…     

镉对集胞藻PCC6803生长的影响

以集胞藻为研究材料,研究不同浓度的Cd2 处理对集胞藻生长的影响.研究发现:低浓度的Cd2 处理(0~0.1 m g/L),能够促进集胞藻的生长,表现为叶绿素、可溶性糖、蛋白质含量随着浓度的增大而增大;当Cd2 浓度进一步增加时,Cd2 开始抑制集胞藻的生长,表现为叶绿素、可溶性糖、蛋白质含量减小,Cd2 浓度达到1.0 m g/L则完全抑制集胞藻的生长,无任何产物累积.     

Effect of Cl- on photosynthetic bicarbonate uptake in two cyanobacteria Microcystis aeruginosa and Synechocystis PCC6803

Photosynthetic inorganic carbon utilization was investigated in two cyanobacteria Microcystis aeruginosa and Synechocystis PCC6803 grown in standing culture. Photosynthetic rates for the two algae reached about 10 times the theoretical CO₂ supply rate at low dissolved inorganic carbon (DIC) of 100 μmol/L, and the rates were unaffected by the addition of 20 mmol/L Na⁺, indicating that the two algae possessed Na+-independent HCO^-₃ utilization for photosynthesis under low DIC. Their photo- synthetic rates at low DIC were inhibited by higher Cl^- and the degrees of inhibition were increased with the rise of CI^- concentration, and in the presence of Diphenylamine-2-carboxylate (DPC), a reported Cl^- channel inhibitor, the rates decreased by 74%－82%, implying that putative DPC-sensitive Cl^- channels participate in Na+-independent HCO₃^- uptake for photosynthesis. The experiment of intracellular 14C-DIC accumulation for photosynthesis showed that internal DIC pools decreased by about 80% with 200 μmol/L DPC and by 64%－70% with 100 mmol/L Cl^-. The experiment of chlorophyll a fluorescence quenching showed that initial rates and extents of fluorescence quenching obviously decreased with 200 μmol/L DPC or 100 mmol/L Cl^-. The two experiments gave further evidence that putative DPC-sen- sitive Cl^- channels participate in Na+-independent HCO^-₃ uptake for photosynthesis in the two algae grown in standing culture.     

Deletion of an electron donor-binding subunit of the NDH-1 complex,NdhS, results in a heat-sensitive growth phenotype in Synechocystis sp. PCC 6803

In cyanobacteria and higher plants, NdhS is suggested to be an electron donor-binding subunit of NADPH dehydrogenase （NDH-1） complexes and its absence impairs NDH-l-dependent cyclic electron trans- port around photosystem I （NDH-CET）. Despite significant advances in the study of NdhS during recent years, its functional role in resisting heat stress is poorly understood. Here, our results revealed that the absence of NdhS resulted in a serious heat-sensitive growth phenotype in the uni- cellular cyanobacterium Synechocystis sp. strain PCC 6803. Furthermore, the rapid and significant increase in NDH-CET caused by heat treatment was completely abolished, and the repair of photosystem II under heat stress conditions was greatly impaired when compared to that of other photosynthetic apparatus in the thylakoid membrane. We therefore conclude that NdhS plays an important role in resistance to heat stress, possibly by stabilizing the electron input module of cyanobacterial NDH-1 complexes.     

封闭式光生物反应器集胞藻6803光自养和混合营养培养比较

采用封闭式光生物反应器进行了蓝藻基因工程常用宿主系统集胞藻Synechocystis sp. PCC 6803的混合营养培养,并与光自养培养进行了比较,在两种培养方式下,集胞藻6803的饱和光强基本相同,都为5000lx,当入射光强为5000lx,初始葡萄糖浓度为1．74g/L时,混合营养生长在葡萄糖消耗完（69．5h)时的藻细胞密度为1．36g/L,叶绿素浓度为20．08mg/L,能量得率为16．7％,分别为同期光自养生长的3．8倍,2．3倍和2．6倍,这表明封闭式光生物反应器混合营养培养方式在促进集胞藻6803生长和光合色素合成及提高培养过程能量得率等方面都有显著作用。     

封闭式光生物反应器集胞藻6803光自养和

采用封闭式光生物反应器进行了蓝藻基因工程常用宿主系统集胞藻Synechocystissp.PCC6803的混合营养培养,并与光自养培养进行了比较.在两种培养方式下,集胞藻6803的饱和光强基本相同,都为5000lx.当入射光强为5000lx、初始葡萄糖浓度为1.74g/L时,混合营养生长在葡萄糖消耗完(69.5h)时的藻细胞密度为1.36g/L、叶绿素浓度为20.08mg/L、能量得率为16.7%,分别为同期光自养生长的3.8倍、2.3倍和2.6倍.这表明封闭式光生物反应器混合营养培养方式在促进集胞藻6803生长和光合色素合成及提高培养过程能量得率等方面都有显著作用.     

绿色荧光蛋白标记S2P在集胞藻6803中的表达

为研究集胞藻6803中两个S2P同源蛋白酶Slr0643和Sll0862的功能,分别在两个蛋白酶的羧基端添加增强型绿色荧光蛋白(EGFP)标签,构建了两株转基因集胞藻psbA2::0643GFP-Cmr/△0643-Kmr和psbA2::0862GFP-Cmr/△0862-Kmr.通过逆转录聚合酶链式反应(RT-PCR)检测到egfp基因的转录表达,通过激光共聚焦显微镜检测到转基因集胞藻细胞发出的绿色荧光蛋白荧光,表明带EGFP标签的S2P融合蛋白在psbA2启动子调控下正确表达.     

Transient Effects of Salt Shock on the Photosynthetic Machinery in Synechocystis sp. PCC 6803

IntroductionSalt stress is one of the main detrimental factors inthe environment that limit the growth andproductivity of plants.Salt stress causessignificant decreases in photosynthetic activity,such as the electron transport[1,2 ] ,but themechanisms by which salt stress inhibitsphotosynthesis remain poorly understood[3] .　Cyanobacteria are prokaryotes that performoxygenic photosynthesis using a photosyntheticapparatus similar to that in the chloroplasts ofgreen algae and higher plants[4 ] .…     

DnaJ-like protein gene sll1384 is involved in phototaxis in Synechocystis sp. PCC 6803

In Synechocystis sp. PCC 6803, gene sll1384 encodes a protein with a DnaJ domain at its N-terminal portion and a TPR domain at the C-terminal portion. An sll1384 mutant shows no difference from the wild type in adaptation to different temperatures, but almost completely loses its capability of phototactic movement. After complementation with sll1384, the mutant regains the phototaxis. As shown with electron microscopy, on the cell surface, mutant cells have pill that appear to be the same as that of the wild type. Also, the transformation efficiency remains unchanged in the mutant. It is postulated that Sll1384 regulates phototaxis of Synechocystis through protein-protein interaction. It is the first DnaJ-like protein gene identified in a cyanobacterium for a role in phototaxis.     

集胞藻胞外多糖的流变学性质和乳化活性

集胞藻在培养过程中向培养基中分泌大量胞外多糖,流变学研究表明这种胞外多糖呈现假塑性行为,粘度值接近黄原胶,当剪切速率在0.15 S-1时,两者粘度相等.集胞藻胞外多糖在0.1 mol/L氯化钠溶液中以及在30～70℃温度范围内粘度值具有一定的稳定性.集胞藻胞外多糖的粘弹谱类似于黄原胶.具有弱凝胶的特征.这种多糖具有良好稳定的乳化活性,对植物油、正十六烷、石蜡、二甲苯和2-甲基萘 正十六烷(1:1)都有乳化作用,而且在静置一个月后,乳液依然稳定.集胞藻胞外多糖可以作为增稠剂、稳定剂以及乳化剂在食品、化妆品工业等领域得到广泛应用.     

温度和集胞藻浓度对轮虫生活史参数的影响

以萼花臂尾轮虫为研究对象,设置3种温度,4个集胞藻浓度梯度,利用动态生命表法探讨了温度和集胞藻浓度对轮虫生活史参数的影响.结果表明,温度、集胞藻浓度以及两者的交互作用均对萼花臂尾轮虫种群的净生殖率(R_0)、世代时间(T)和内禀增长率(r_m)有极显著影响.温度为15℃时,轮虫的净生殖率随集胞藻浓度的升高而显著增加,温度为25℃和35℃时,轮虫的净生殖率随集胞藻浓度的升高出现下降的趋势.随温度的升高,轮虫世代时间明显缩短;相同温度下,不同实验组轮虫的世代时间也随集胞藻浓度的升高而趋于缩短.当温度为15℃时,轮虫的内禀增长率随集胞藻浓度的升高而显著增加,但是随着温度升高,集胞藻对于轮虫内禀增长率的促进作用减弱.结果显示,集胞藻可以为轮虫种群提供一定的营养,温度的升高降低了集胞藻对轮虫种群的促进作用,本研究结果为温度变化下浮游动物种群对不同蓝藻种类的响应提供了数据基础.     

补糖与运动后糖原合成

论述了补糖时间对补糖数量及种类对运动员肝、肌糖原合成的影响，并对肌肉损伤与糖原合成、运动后补糖与运动能力的关系进行了讨论     

Changes in the transthylakoid proton gradient are caused by the movement of phycobilisomes in the cyanobacterium <Emphasis Type="Italic">Synechocystis</Emphasis> sp. strain PCC 6803

Phycobilisomes (PBSs) are the main accessory light-harvesting complexes in cyanobacteria and their movement between photosystems (PSs) affects cyclic and respiratory electron transport. However, it remains unclear whether the movement of PBSs between PSs also affects the transthylakoid proton gradient (ΔpH). We investigated the effect of PBS movement on ΔpH levels in a unicellular cyanobacterium Synechocystis sp. strain PCC 6803, using glycinebetaine to immobilize and couple PBSs to photosystem II (PSII) or photosystem I (PSI) by applying under far-red or green light, respectively. The immobilization of PBSs at PSII inhibited decreases in ΔpH, as reflected by the slow phase of millisecond-delayed light emission (ms-DLE) that occurs during the movement of PBSs from PSII to PSI. By contrast, the immobilization of PBSs at PSI inhibited the increase in ΔpH that occurs when PBSs move from PSI to PSII. Comparison of the changes in ΔpH and electron transport caused by the movement of PBSs between PSs indicated that the changes in ΔpH were most likely caused by respiratory electron transport. This will further improve our understanding of the physiological role of PBS movement in cyanobacteria.     

Bifunctional enzyme FBPase/SBPase is essential for photoautotrophic growth in cyanobacterium Synechocystis sp. PCC 6803

From a random insertion mutant library of Synechocystis sp. PCC 6803, a mutant defective in photoautotrophic growth was obtained. The interrupted gene was identified to be slr2094 (fbp1), which encodes the fructose-1,6-biphosphatase (FBPase) / sedoheptulose-1,7-biphosphatase (SBPase) bifunctional enzyme (F-I). Two other independently constructed slr2094 mutants showed an identical phenotype. The FBPase activity was found to be virtually lacked in slr2094 mutant, which was sensitive to light under mixotrophic growth conditions. These results indicate that slr2094 is the only active FBPase-encoding gene in this cyanobacterium. Inactivation of photosystem II by interrupting psbB in slr2094 mutant alleviated the sensitiveness to light. This report provides the direct genetic evidence for the essential role of F-I in the photosynthesis of Synechocystis sp. PCC 6803.