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101.
【目的】本研究旨在克隆来自北极海洋、具有合成卤化物潜力的链霉菌(Streptomyces sp.)604F中的一个卤化酶基因,为后续克隆卤化物合成基因簇、分离鉴定卤化物提供指导。【方法】利用琼脂块法初步测试抗菌活性,借助液相色谱-飞行时间串联质谱技术(LC-Tof MS)初步寻找Streptomyces sp.604F发酵粗提液中的卤化物,并以简并PCR扩增合成次级代谢产物的指示基因(I型、II型聚酮合酶及非核糖体多肽合成酶编码基因);根据依赖黄素腺嘌呤二核苷酸的卤化酶基因保守区设计的简并引物,扩增基因片段并测序分析;采用高效热不对称交错PCR(hiTAIL-PCR)技术扩增卤化酶基因全长。【结果】Streptomyces sp.604F具有较强的抗白色念珠菌活性,其基因组同时含有编码I型聚酮合酶、II型聚酮合酶和非核糖体多肽合成酶基因,以及卤化酶基因;通过染色体步移克隆该卤化酶基因全长,共1443 bp,编码一个新的非色氨酸卤化酶,在合成已知卤化物的卤化酶数据库中,与其同源关系最近的为一类参与合成糖肽类次级代谢产物的卤化酶。【结论】Streptomyces sp.604F具有新的非色氨酸卤化酶,且推测参与糖肽类化合物的卤化修饰,为后续寻找目的卤化物提供了指导,也为研究该合成基因簇奠定基础。  相似文献   
102.
Cell fate commitment of pre-implantation blastocysts, to either the inner cell mass or trophoblast, is the first step in cell lineage segregation of the developing human embryo. However, the intercellular signals that control fate determination of these cells remain obscure. Human embryonic stem cells (hESCs) provide a unique model for studying human early embryonic development. We have previously shown that Activin/Nodal signaling contributes to maintaining pluripotency of hESCs, which are derivatives of the inner cell mass. Here we further demonstrate that the inhibition of Activin/Nodal signaling results in the loss of hESC pluripotency and trophoblast differentiation, similar to BMP4-induced trophoblast differentiation from hESCs. We also show that the trophoblast induction effect of BMP4 correlates with and depends on the inhibition of Activin/Nodal signaling. However, the activation of BMP signaling is still required for trophoblast differentiation when Activin/Nodal signaling is inhibited. These data reveal that the early lineage segregation of hESCs is determined by the combinatorial signals of Activin/Nodal and BMP.  相似文献   
103.
Experiments were conducted to determine whether sucrose synthase (SuSy) was phosphorylated in the elongation zone of maize (Zea mays L.) leaves. The approximately 90-kD subunit of SuSy was 32P-labeled on seryl residue(s) when excised shoots were fed [32P]orthophosphate. Both isoforms of SuSy (the SS1 and SS2 proteins) were phosphorylated in vivo, and tryptic peptide-mapping analysis suggested a single, similar phosphorylation site in both proteins. A combination of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and automated Edman sequencing analysis unequivocally identified the phosphorylation site in the maize SS2 protein as serine-15. This site was phosphorylated in vitro by endogenous protein kinase(s) in a strictly Ca(2+)-dependent manner. A synthetic peptide, based on the phosphorylation site sequence, was used to identify and partially purify an endogenous Ca(2+)-dependent protein kinase(s) from the maize leaf elongation zone and expanding spinach leaves. Phosphorylation of SuSy in vitro selectively activates the cleavage reaction by increasing the apparent affinity of the enzyme for sucrose and UDP, suggesting that phosphorylation may be of regulatory significance. Conservation of the phosphorylation site, and the sequences surrounding it, among plant species suggests that phosphorylation of SuSy may be widespread, if not universal, in plants.  相似文献   
104.
105.
应用紫外诱变技术对溶藻菌株NP23进行紫外诱变处理。经过粗筛后,从8株诱变株中选出2株对绿藻中小球藻和蓝藻中惠氏微囊藻的去除效果明显优于原始菌株的突变株NP23-1和NP23-4,其溶藻率(叶绿素a的去除率)比原始菌株提高30%-35%。连续6代测试,2株诱变菌株NP23-1和NP23-4溶藻率都很稳定,表明所得突变株是比原始菌株更优秀的溶藻菌株。  相似文献   
106.
Liao Y  Chen G Z 《农业工程》2007,27(6):2208-2214
The impact of salinity on three arboreal mangrove plants, Sonneratia apetala (Sa), S. caseolaris (Sc) and Rhizophora stylosa (Rs), was studied. The three mangrove species were treated with different salinity levels over a three-month period. The response and adaptation of these three mangrove species to salinity were shown to be different. Net photosynthesis rate, stomata conductance and transpiration rate of leaves decreased and soluble sugar content in leaves increased, with salt concentration in all three mangrove species. The malondial dehyde (MDA) content in stems and leaves of Sa and Sc somewhat decreased when the salinity was lower than 10, but rapidly increased with increasing salt concentration. The MDA content in stems and leaves of Rs increased only when salinity was greater than 40. No changes were observed in the MDA content of roots in the three mangrove species. The adaptabilities of Sa and Sc to salt tolerance were limited. The more salt tolerant the mangrove Rs, the more likely the free oxygen radicals were eliminated through the increase in activity of superoxide dismutase (SOD). Results of this experiment identified salinity levels best suited for the growth and metabolism of the species, which provides information necessary for maintaining mangrove forestation along the South China coast.  相似文献   
107.
Wu J  Du H  Liao X  Zhao Y  Li L  Yang L 《Plant molecular biology》2011,77(1-2):117-127
We have developed a modified particle bombardment method for plant transgenesis. An intein-tag and a 6×Cys-tag were successively fused to the N-terminus of a hyperactive Tn5 transposase. The modified transposase was immobilized on bare gold microscopic particles via covalent binding of a 6×Cys-tag sulfydryl groups to the gold surface. The tethered transposase can bind the transposon DNA in vitro to form the transposome in the absence of Mg2? ions. After bombardment of the gold particles carrying the transposomes into the plant cells, the transposomes will be released from the carrier due to the activated self-cleavage function of intein-tag. Our data showed this procedure integrated foreign DNA into the plant genome with an increased transformation frequency as compared to the conventional particle bombardment method. A single copy insertion can also be obtained by decreasing of the assembled transposon DNA amount in relation to plant cell biomass.  相似文献   
108.
Shen CI  Wang CH  Shen SC  Lee HC  Liao JW  Su HL 《PloS one》2011,6(5):e18894
Sialic acids (SAs) linked to galactose (Gal) in α2,3- and α2,6-configurations are the receptors for avian and human influenza viruses, respectively. We demonstrate that chicken tracheal ciliated cells express α2,3-linked SA, while goblet cells mainly express α2,6-linked SA. In addition, the plant lectin MAL-II, but not MAA/MAL-I, is bound to the surface of goblet cells, suggesting that SA2,3-linked oligosaccharides with Galβ1-3GalNAc subterminal residues are specifically present on the goblet cells. Moreover, both α2,3- and α2,6-linked SAs are detected on single tracheal basal cells. At a low multiplicity of infection (MOI) avian influenza virus H6N1 is exclusively detected in the ciliated cells, suggesting that the ciliated cell is the major target cell of the H6N1 virus. At a MOI of 1, ciliated, goblet and basal cells are all permissive to the AIV infection. This result clearly elucidates the receptor distribution for the avian influenza virus among chicken tracheal epithelial cells and illustrates a primary cell model for evaluating the cell tropisms of respiratory viruses in poultry.  相似文献   
109.
110.
Leaf mass per unit area (LMA), carbon and nitrogen contents, leaf construction cost, and photosynthetic capacity (P max) of Adiantum reniforme var. sinensis, an endangered fern endemic to the Three Gorges region in southwest China, were compared in five populations differing in habitat such as soil moisture and irradiance. The low soil moisture and high irradiance habitat population exhibited significantly higher LMA, area-based leaf construction (CCA), and carbon content (CA), but lower leaf nitrogen content per unit dry mass (NM) than the other habitat populations. The high soil moisture and low irradiance habitat populations had the lowest CCA, but their cost/benefic ratios of CCA/P max were similar to the medium soil moisture and irradiance habitat population due to their lower leaf P max. Hence A. reniforme var. sinensis prefers partially shaded, moist but well-drained, slope habitats. Due to human activities, however, its main habitats now are cliffs or steeply sloped bare rocks with poor and thin soil. The relatively high energy requirements and low photosynthetic capacity in these habitats could limit the capability of the species in extending population or interspecific competition and hence increase its endangerment.  相似文献   
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