Paired cloning vectors for complementation of mutations in the cyanobacterium<Emphasis Type="Italic"> Anabaena</Emphasis> sp. strain PCC 7120

The clones generated in a sequencing project represent a resource for subsequent analysis of the organism whose genome has been sequenced. We describe an interrelated group of cloning vectors that either integrate into the genome or replicate, and that enhance the utility, for developmental and other studies, of the clones used to determine the genomic sequence of the cyanobacterium, Anabaena sp. strain PCC 7120. One integrating vector is a mobilizable BAC vector that was used both to generate bridging clones and to complement transposon mutations. Upon addition of a cassette that permits mobilization and selection, pUC-based sequencing clones can also integrate into the genome and thereupon complement transposon mutations. The replicating vectors are based on cyanobacterial plasmid pDU1, whose sequence we report, and on broad-host-range plasmid RSF1010. The RSF1010- and pDU1-based vectors provide the opportunity to express different genes from either cell-type-specific or -generalist promoters, simultaneously from different plasmids in the same cyanobacterial cells. We show that pDU1 ORF4 and its upstream region play an essential role in the replication and copy number of pDU1, and that ORFs alr2887 and alr3546 (hetF _A ) of Anabaena sp. are required specifically for fixation of dinitrogen under oxic conditions.     

An inducible system for expression and validation of the specificity of short hairpin RNA in mammalian cells

RNA interference (RNAi) by means of short hairpin RNA (shRNA) has developed into a powerful tool for loss-of-function analysis in mammalian cells. The principal problem in RNAi experiments is off-target effects, and the most vigorous demonstration of the specificity of shRNA is the rescue of the RNAi effects with a shRNA-resistant target gene. This presents its own problems, including the unpredictable relative expression of shRNA and rescue cDNA in individual cells, and the difficulty in generating stable cell lines. In this report, we evaluated the plausibility of combining the expression of shRNA and rescue cDNA in the same vector. In addition to facilitate the validation of shRNA specificity, this system also considerably simplifies the generation of shRNA-expressing cell lines. Since the compensatory cDNA is under the control of an inducible promoter, stable shRNA-expressing cells can be generated before the knockdown phenotypes are studied by conditionally turning off the rescue protein. Conversely, the rescue protein can be activated after the endogenous protein is completely repressed. This approach is particularly suitable when prolonged expression of either the shRNA or the compensatory cDNA is detrimental to cell growth. This system allows a convenient one-step validation of shRNA and generation of stable shRNA-expressing cells.     

Nickel-Resistant Determinant from Leptospirillum ferriphilum

Leptospirillum ferriphilum strain UBK03 isolated from a mine in Jiangxi, China, is resistant to Ni²⁺ (30 to 40 mM). A four-gene nickel resistance cluster was identified and, when transformed into Escherichia coli, enabled growth in 6 mM nickel. Mutation experiments revealed that the genes ncrA, ncrB, and ncrC could confer nickel resistance in Escherichia coli, whereas the gene ncrY could have a negative effect on nickel resistance.     

Removal of agricultural non-point source pollutants by ditch wetlands: implications for lake eutrophication control

Ditches grown with nature reed (Phragmites communis Trin) and wild rice (Zizania latifolia Turcz) were selected to study the removal capacity of agricultural non-point source pollutants so as to find a way to alleviate eutrophication in Lake Taihu. Ditches sediment from depths below 40 cm can accumulate organic matter and total nitrogen (TN). TN is correlated positively to organic matter in reed populated sediment and wild rice populated sediment. This suggests that the main composition of TN is organic nitrogen derived from plant decomposition. A significant negative relationship between TN and pH was found in reed and wild rice sediments. Seasonal harvest of helophyte vegetation is an effective method to remove N and P from wetlands. Organic matter and TN concentrations in water and sediments (0–20 cm) in areas where reeds were removed are lower than non-harvested areas (control). Reeds and wild rice have high uptake ability of nitrogen (N) and phosphorus (P). However, the low economic value of these plants will not stimulate voluntary harvest of farmers. Zizania caduciflora Turez Hand-mazt is a kind of vegetable widely cultivated in ditches around the lake. It can also absorb N and P effectively. Thus, large scale cultivation of Z. caduciflora to replace nature plants may improve water quality.     

Homochiral metal-organic coordination networks from l-typtophan

Three homochiral metal-organic coordination networks [Co₂(l-Trp)₂(Py)₆] · Py · (ClO₄)₂ (1), [Ni(l-Trp)(Py)₃] · H₂O · ClO₄ (2) and [Co₂(l-Trp)(INT)₂(H₂O)₂(ClO₄)] (3), all containing natural amino acid l-HTrp (l-typtophan), were hydrothermally synthesized and structurally characterized. The compounds 1 and 2 crystallize in the orthorhombic space group C222₁, with a = 10.731(2) Å, b = 19.709(4) Å, c = 27.365(6) Å and Z = 4 for 1 and a = 10.710(10) Å, b = 20.088(18) Å, c = 27.63(3) Å and Z = 8 for 2, respectively. The compound 3 has the monoclinic space group P2₁, with a = 8.1934(14) Å, b = 13.209(2) Å, c = 12.464(2) Å, β = 104.107(3)° and Z = 2. Both 1 and 2 consist of 1D helical chains. Compound 3 is composed of 2D networks, which further assemble into a 3D supramolecular structure via weak interlayer interactions. The optically pure amino acid l-HTrp plays an important role leading to homochiral structures reported here.     

Predicted glycosyl transferase genes located outside the HEP island are required for formation of heterocyst envelope polysaccharide in Anabaena sp. strain PCC 7120

During maturation, heterocysts form an envelope layer of polysaccharide, called heterocyst envelope polysaccharide (HEP), whose synthesis depends on a cluster of genes, the HEP island, and on an additional, distant gene, hepB, or a gene immediately downstream from hepB. We show that HEP formation depends upon the predicted glycosyl transferase genes all4160 at a third locus and alr3699, which is adjacent to hepB and is cotranscribed with it. Mutations in the histidine kinase genes hepN and hepK appear to silence the promoter of hepB and incompletely down-regulate all4160.     

云南省小黑山自然保护区兰科植物多样性及保护评价

云南省小黑山自然保护区孕育了丰富的兰科植物。据调查,保护区有兰科植物43属、134种,约占云南省135属、780种的31.9%和17.2%,是保护区种子植物种类最多的科。兰科植物在保护区的各个生态类型中均有出现。保护区134种兰科植物以附生兰占优势,占67.16%(90种);其次,地生兰占31.34%(42种),腐生兰为1.50%(2种)。保护区兰科植物起源于新、旧世界的热带和温带,热带属占70.0%(28),温带属占27.5%(11)。鉴于兰科植物重要的保护价值,应加强保护区兰科植物的保护。首先应加强保护区的建设和管理,还应加强社区经济发展,加强科学研究,实施社区共管以及加大保护宣传力度。     

大鼠三叉神经节不同直径神经元ATP-激活电流的特征

目的:探讨大鼠三叉神经节不同直径神经元ATP-激活电流的特征。方法:应用全细胞膜片钳技术进行实验。结果:①92.3%(60/65)的细胞对ATP敏感,有反应的细胞可记录到三种型式的ATP-激活电流:快速激活快速失活型(Fast type,F型)、快速激活缓慢失活型(Intermediate type,I型)和缓慢激活缓慢失活型(Slowtype,S型)。三种电流均具有浓度依赖性。②小直径的细胞多表现为F型特征,大直径的细胞多表现为S型特征,而中等大小的细胞多表现为I型特征。③动力学特征:三种类型的ATP激活电流上升相从10%到90%的时间:F型:(33.6±4.5)ms;Ⅰ型:(62.2±9.9)ms;S型:(302.1±62)ms。去敏感相从10%到90%的时间:F型:(399.4±58.2)ms;S型:>500ms。④I-V曲线:三种电流均表现为内向整流的特性,而且翻转电位均为0~5mV。⑤量-效关系:Ⅰ型的量-效曲线居中间,F型的下移,S型的上移,三种类型电流量-效曲线的EC50非常接近。结论:三种型式的ATP-激活电流可能是由不同亚单位组合的P2X受体各亚型所介导,这些亚型分布于不同大小的三叉神经节神经元,从而传导不同的信息。     

Deficiency of DNA fragmentation factor 45 results in reduced oocyte apoptosis in response to doxorubicin

Dear Editor： Apoptosis plays a prominent role in ovarian development and function. During follicle development, the vast majority of follicles undergo atresia as a consequence of apoptosis of constituent oocyte or follicular cells, or both, failing to complete the maturation process. Atresia occurs at all stages of follicular development during the growth and development of follicles.[第一段]