Cloning and analysis of [NiFe]-hydrogenase genes from Arthrospira platensis FACHB341

0IntroductionHydrogenase(H2ase)has beeninvestigatedin a va-riety of bacterial groups since it was firstly reported in1931[1-3].Cyanobacteria contain two different types ofH2ase,uptake H2ase(EC1.12.7.2)and bidirectional orreversible H2ase(EC1.12.1.12).Uptake H2ase is in-duced under N2-fixing situation,mainly confinedin hete-rocysts.Bidirectional H2ase is constitutively synthesized,active in both heterocysts and vegetative cells present inboth N2-fixing and non-N2-fixing conditions.This en…     

极大节旋藻气囊蛋白基因gvpA克隆与序列分析

为了研究极大节旋藻气囊蛋白的分子生物学特性,并为gvpA基因的研究提供基础材料,在对极大节旋藻GSS序列分析的基础上,采用PCR技术克隆了gvpA基因的全长序列,并进行了相关分析。结果表明极大节旋藻gvpA基因编码区的GC含量为42.92%,核苷酸序列和蛋白序列与Planktothrix agardhii的gvpA基因相似性最高(79.4%,97.2%),其次是Pseudanabaena sp.PCC6901(75.3%,94.4%)。系统发生分析表明极大节旋藻gvpA基因与Planktothrix agardhii的gvpA基因同源性最高(89%ML,85%MP,89%NJ)。     

Characteristics of rDNA intergenic spacer region from a waterbloom cyanobacteria species Oscillatoria sp.

Sequence of rDNA intergenic spacer region (ISR) from a waterbloom cyanobacterial species Oscilla-toria sp. was determined and analyzed. The results of sequence comparison showed that the spacer had a high level sequence divergence, suggesting the sequence may be a target sequence for developing cyanobacteria genus- and species-specific oligonucleotide probes. In addition, a 20bp sequence of rDNA ISR was found highly conserved in all species of cyanobacteria, which was not found in other eubacteria. This conserved sequence within a variable region indicates that it might be a functional oligonucleotide in the processing of the rRNA precursor.     

β-胡萝卜素原料筛选及其超临界二氧化碳萃取

建立了柱层析／分光光度计法测定β－胡萝卜素的分析方法。筛选出富含β－胡萝卜素的螺旋藻做为原料。分别以石油醚和超临界二氧化碳萃取螺旋藻中β－胡萝卜素。结果表明,超临界二氧化碳萃取藻粉中β－胡萝卜素与石油醚萃取相比具有萃取效率高、速度快、工艺简单、萃取物色味纯正、藻粉可以进一步利用和再出售的优点。     

氯化胆碱在钝顶螺旋藻(Spirulina platensis)养殖中的应用初探

螺旋藻是联合国粮农组织和联合国工业发展组织推荐的营养保健品,它的丰富营养物质和明显的药效作用已引起国际上的广泛重视,被誉为明天最理想的保健品和超级食品［１,２］．近来螺旋藻产业在国内外形成了一定的规模,但也存在着生产成本高的问题,如何降低生产成本已成...     

A miniature insertion element transposable in Microcystis sp. FACHB 854

A 186-bp sequence with imperfect terminal inverted repeats and target direct repeats but without any transposase-encoding capacity was found to be transposable in an isolate derived from Microcystis sp. FACHB 854. This miniature insertion element, designated as ISM854-1, and with its homologues present at least 10 copies in the genome of Microcystis FACHB 854, is inserted into the 8-bp long and AT-rich target sequences, but none or few in other Microcystis strains. A variant of ISM854-1, denoted ISM854-1A, has perfect inverted repeat sequences and may transpose in pairs in a structure like a composite transposon. This is the first report of non-autonomous transposition of a mini-IS in a cyanobacterium.     

仔猪先天性震颤的病原学调查及其病原特性分析

应用PCR方法对临床收集的8个猪场仔猪先天性震颤病例8例进行CSFV、PCV-2和PPV检测,结果 CSFV阳性率为87.5%,PCV-2阳性率为87.5%,PPV阳性率为37.5%。其中CSFV、PCV-2和PPV混合感染率为25.0%,CSFV和PCV-2混合感染率为50.0%,CSFV和PPV混合感染率为12.5%。序列分析结果显示:7株CSFV的E2基因之间核苷酸序列差异很小,同源性为99.0%～100%,与Alfort株同源性为95.1%～96.2%,与猪瘟兔化弱毒株(HCLV)和石门株(Shimen)的同源性为83.3%～84.7%。5株PCV-2 Cap基因之间核苷酸序列差异很小,分离的同源性为98.9%～100%,与Genbank发布的标准序列的同源性为98.7%～99.4%;3株PPV VP2基因之间核苷酸序列差异也很小,同源性为99.4%～99.6%,与Genbank发布的标准序列的同源性为99.0%～99.8%。PRV动物接种试验结果显示:8个病例的病料组织悬液接种家兔,家兔表现正常,PRV野毒感染试验阴性。     

Vacuolation induced by unfavorable pH in cyanobacteria

Six species or strains of cyanobacteria, Anabaena sp. 595, Plectonema boryanum 246, Scytonema hofmanni 248, Nostoc sp. 96, Oscillatoria animlis 284 and Spirulina maxima 438, were cultured in unfavorable pH conditions for vacuole induction. At pH 5.0, 6.5, or 7.0, vacuoles were observed to form in both Anabaena sp. 595 and Plectonema boryanum 246, especially in the former. The vacuolation took place with some morphological changes, such as the cells being inflated, spherical and vacuolated, and with unequalized division. The induced vacuoles in An- abaena sp. 595 and Plectonema boryanum 246 were in spherical shape and in rather transparent appearance under a phase microscope. For Scytonema hofmanni 248, it was less sensitive to pH, its vacuole formation was found only at pH 6.5. No vacuolization occurred in the cells of Nostoc sp. 96, Oscillatoria animlis 284 and Spirulina maxima 438 at all pH conditions we used. Vacuolization under unfavorable pH provides a new proof for the existence of vacuole in cells of cyanobacteria and reflects the prokaryote's function in ecological environment.     

蓝藻基因表达载体系统的构建和应用

蓝藻是一类具植物型放氧光合作物特性的原核生物。多数蓝藻富含营养物质,无毒,是表达外源目的基因的独特受体系统。因此,构建适用于蓝藻表达外源目的基因的载体系统,并用于表达药物活性肽基因,已成为当前蓝藻基因工程研究的热点之一。本课题组10多年来率先在国内开展蓝藻基因工程研究,从蓝藻Plectonema botyanum中分离得到了约1.5kb的小质粒,以此为出发质粒,构建了蓝藻穿梭质粒pPRS-1和穿梭质粒表达载体pPKE2;同时根据DNA片段同源重组的性质,构建了蓝藻Calothrix sp.PCC7601、Synechococcus sp.PCC7942的基因整合平台系统。在国家863项目经费资助下,利用构建的蓝藻质粒载体和基因整合平台系统,把人源胸腺素基因转入蓝藻Calothrix sp.PCC7601和Synechoccus sp.PCC7942,并能高效表达,首次获得了可直接口服的含人源胸腺素的转基因蓝藻,这对研究和开发基因工程口服药物具有重大的科学意义和经济、社会效益。     

外源AHLs信号物质对蓝藻生长代谢的影响

 研究了外加AHLs(acyl-HSLs)信号物质对滇池铜绿微囊藻(Microcystis aeruginosa)FACHB905以及集胞藻(Synechocysti sp.)FACHB898生长和代谢的影响.结果表明:AHLs类信号物质对2种藻具有明显的生长抑制作用.在10~80μmol·L^-1浓度范围内,2种供试藻的生物量,藻蓝蛋白(phycocyanin,PC)含量都受到了一定程度的抑制,而2种藻的可溶性糖含量和Car/Chl a却有明显地提高.随AHLs浓度的增加,铜绿微囊藻藻液中超氧自由基含量明显地增加,而集胞藻藻液中超氧自由基含量随AHLs物质浓度的增加呈下降趋势,反映出两者在适应性上是有差异的.     

Kas基因启动子区的克隆及功能初步分析

利用TAIL-PCR技术,克隆到了与辣椒素合成有关的胎座特异表达基因——3-酮酯酰．ACP合成酶基因（Kas）上游400bp的调控区域．将其全长片段与GUS基因连接构建植物表达载体并转化烟草．GUS组织化学染色表明,克隆到的440bp片段具有启动子活性．对该片段进行序列分析发现,在起始密码子ATG上游存在2个TATA-box,分别为-316～-311位的TATAAA和-224～-219位的TATAAA;在TATA-box上游还存在1个位于-378～-374处的CAAT-box,序列为CCAAT．该研究旨在为利用基因调控辣椒素的生物合成,提高辣椒果实中的辣椒素含量奠定基础．     

玫瑰微球菌海藻糖合成相关酶基因的克隆和序列分析

从玫瑰微球菌QS412中克隆出海藻糖生成相关酶――麦芽寡糖基海藻糖基水解酶的基因treZ ,测定了其核苷酸序列并进行了表达。treZ 编码的蛋白质有624个氨基酸、分子质量为68 kD. 它们与已报道的其他微生物的海藻糖生成相关酶的基因进行同源性比较，treZ 的同源性分别为33.0%(耐放射异常球菌)；10.1%(硫矿硫化叶菌KM1)；51.9%(节杆菌Q36)；52.8%(根瘤菌M11)；48.5% (短杆菌)。经过氨基酸序列比较分析还发现，所有的海藻糖生成相关酶都含有糖苷酶家族13个中几个高度保守的α-淀粉酶催化活性区，推测这些海藻糖生成相关酶都可能有着共同的进化来源。     

关于不定方程χ~3+1=201y~2

利用Pell方程、递归数列的方法证明了不定方程χ~3+1=201y~2的整数解只有(-1,0),(440,651),(440,-651).     

Hydrogen ions directly regulating the oligomerization state of Photosystem I in intact <Emphasis Type="Italic">Spirulina platensis</Emphasis> cells

H^＋ concentration induced-monomerization or trimerization of photosystem Ⅰ （PSI） in cyanobacteria has never been directly observed. In this work, taking characteristic spectra for the trimers and monomers as the indicators, it was experimentally demonstrated that H^＋ could induce the oligomeric changes of PSI reaction centers in the intact Spirulina platensis cells and also in the isolated thylakoid membrane complexes. Especially, the higher concentration of H^＋ would induce the monomerization while the lower the trimerization, suggesting the electrostatic interaction should be mainly responsible for changes in the oligomeric state of PSI in Spirulina platensis.     

螺旋藻的磁处理培养

就磁场处理技术对螺旋藻培养过程的强化进行了研究，发现螺旋藻的培养能明显地被适当的磁场处理所刺激，在强度为200－320kA/m的外加磁场作用下培养至第6天，螺旋藻最大细胞干重达2.76g/L,比同等条件下的空白对照试样多46．8％，其比生长率由0.4d^-1增至0.6d^-1，培养周期可缩短2－3d,同时，螺旋藻中蛋白质的含量增加了5.2mg/g，氨基酸总含量（除色氨酸外）增加了0.71mg/g，其中必需氨基酸增加了3.15mg/g，此外，微量元素Sr,Ni,Cu,Mn和Zn等均有显著增加，其中Sr和Ni分别增加了22.3和5．1倍，文中最后讨论了磁场处理对螺旋藻培养的强化机制，指出这种刺激作用与磁场处理加速了螺旋藻的光合作用和强化了其营养吸收有关。     

有机碳氮源对钝顶螺旋藻生长及叶绿素a含量的影响

在Zarrouk溶液中添加不同的有机碳氮源,研究它们对钝顶螺旋藻生长及叶绿素a含量的影响。结果表明:糖类中,浓度为1.5 g.L-1葡萄糖对钝顶螺旋藻生长影响明显,平均生长速率增加了19.1%,平均世代时间减少了16.0%,其次为麦芽糖,平均生长速率增加了5.2%,平均世代时间减少了5.6%;有机酸钠盐对螺旋藻的生长影响不太明显,只有苹果酸盐对其生长有一定的促进作用。有机氮源中,只有0.2 g.L-1尿素有微弱的促长作用。添加的有机碳氮源中,苹果酸钠对螺旋藻叶绿素a含量的影响最为明显,与对照相比,螺旋藻单位藻体的叶绿素a含量提高了73.4%,葡萄糖则使单位藻体的叶绿素a含量有所下降。     

甜菜夜蛾核多角体病毒（SeMNPV）gp 37基因的克隆及序列分析

通过鸟枪法克隆了甜菜夜蛾核多角体病毒（SeMNPV)基因组DNA EcoR I酶切的2．2kb片段,序列分析表明,该片段含有gp37基因,SeMNPV gp37基因的开放阅读框为801个核苷酸,编码268个氨基酸,预测蛋白质相对分子质量为30400,在gp37基因起始密码子上游有一典型的杆状病毒晚期基因启动子序列ATAAG,同其它昆虫杆状病毒和昆虫痘病毒GP37／Fusolin蛋白的比较结果表明,SeMNPV GP37与已知gp37/fusolin基因的氨基酸序列同源性较高（50－68％）,在其蛋白序列内存在类似的保守区和可能的N－连接糖基化位点,在SeMNPV gp37基因下游存在一个完整阅读框和部分get基因序列。     

Primary structure of helix C to helix G of a new retinal protein in H.sp.xz515

The gene fragment encoding the retinal protein from helix C to helix G in a new strain of halobacteria, H.sp.xz515 has been amplified by PCR method. The nucleotide sequence of this fragment has been determined. The deduced amino acid sequence has been compared with halobium br and other two br-like proteins, ar-1 and ar-2. Results show that those amino acid residues in br, essential for proton pumping and binding to retinal, are conserved. The residue M145 in br may be important for isomerization reaction of retinal.     

淀山湖蓝藻水华暴发的气象水文因素探讨

2007-2009年淀山湖的总氮和总磷年均浓度分别为3.47～4.53 mg/L和0.18～0.24mg/L,明显高于富营养化水平,成为蓝藻水华暴发的重要物质基础.MODIS影像分析结果表明,2007-2009年蓝藻水华暴发频次分别为3,6,10 d,呈逐年上升的趋势,并且暴发后易于集中在湖中心、东北和东部水域.受黄浦...