喀喇昆仑山-西昆仑山地区土壤元素的自然含量特征

本文对喀喇昆仑山-西昆仑山地区土壤中23种元素的自然含量进行了论证。用对比和区域分异系数的方法,讨论了本区土壤元素在世界、我国和区域内的异同和分异特征。通过相关分析和主成份分析,揭示了土壤元素的共生组合关系和表现在土壤类型上的聚类特征,进一步说明了元素在本区土壤中的相关性和分异特征。     

Growth promotion effects of the endophyte <Emphasis Type="Italic">Acinetobacter johnsonii</Emphasis> strain 3-1 on sugar beet

An endophytic bacteriumn identified as Acinetobacter johnsonii strain 3–1 was isolated from surface-sterilized roots of Beta vulgaris. Its effect on sugar beet seedling growth was studied using pot assays and field experiments. This strain promoted beet seedling growth following seed inoculation by seed dipping. Plant height and dry weight of beet increased by 19% and 69%, respectively, compared with controls. Strain 3–1 exhibited the ability to increase absorption of N, P, K, and Mg elements from soil and increase the content of vitamins B and C, and protein within beet. In addition, the strain also produced a phytohormone-auxin, produced nearly twice as much IAA as that produced by strain 2–2, and was able to solubilize phosphates. The concentration of dissolved P in the medium was 180.5 mg L⁻¹ after 4 days of incubation. In field experiments, strain 3–1 significantly increased the content of sucrose, fructose, and the yield of the beet. The growth-promoting properties of Acinetobacter johnsonii strain 3–1 indicates that this promising isolate merits further investigation into its symbiosis with beet plants and its potential application in agriculture.     

Generation of an Oocyte-Specific Cas9 Transgenic Mouse for Genome Editing

The CRISPR/Cas9 system has been developed as an easy-handle and multiplexable approach for engineering eukaryotic genomes by zygote microinjection of Cas9 and sgRNA, while preparing Cas9 for microinjection is laborious and introducing inconsistency into the experiment. Here, we describe a modified strategy for gene targeting through using oocyte-specific Cas9 transgenic mouse. With this mouse line, we successfully achieve precise gene targeting by injection of sgRNAs only into one-cell-stage embryos. Through comprehensive analysis, we also show allele complexity and off-target mutagenesis induced by this strategy is obviously lower than Cas9 mRNA/sgRNA injection. Thus, injection of sgRNAs into oocyte-specific Cas9 transgenic mouse embryo provides a convenient, efficient and reliable approach for mouse genome editing.     

Synthesis of 4/5-deoxy-4/5-nucleobase derivatives of 3-O-methyl-D-chiro-inositol as potential antiviral agents

Using D-pinitol (= 3-O-methyl-D-chiro-inositol) as starting material, a concise synthesis of 4/5-deoxy-4/5-nucleobase derivatives 11-19 has been achieved. The key intermediate 9 was obtained in good yield via an epoxidation from mono-methanesulfonate of D-pinitol. The process of opening of the epoxide ring in 9 by nucleobases appeared to be regioselective in presence of 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). All the synthesized carbocyclic nucleosides were assayed against several viruses and tumors such as HIV-1, HSV-1, and HSV-2, and lung and bladder cancer. However, only compounds 14b, 14a, 16a, 16b, and 19 showed mild inhibitory effect against human lung cancer cell lines (PG) with IC50 values ranging from 50 to 100 microM, and the other compounds did not exhibit any significant antiviral activity or cytotoxicity even at concentrations up to 200 microM.     

A Novel Signal Transduction Pathway that Modulates rhl Quorum Sensing and Bacterial Virulence in Pseudomonas aeruginosa

The rhl quorum-sensing (QS) system plays critical roles in the pathogenesis of P. aeruginosa. However, the regulatory effects that occur directly upstream of the rhl QS system are poorly understood. Here, we show that deletion of gene encoding for the two-component sensor BfmS leads to the activation of its cognate response regulator BfmR, which in turn directly binds to the promoter and decreases the expression of the rhlR gene that encodes the QS regulator RhlR, causing the inhibition of the rhl QS system. In the absence of bfmS, the Acka-Pta pathway can modulate the regulatory activity of BfmR. In addition, BfmS tunes the expression of 202 genes that comprise 3.6% of the P. aeruginosa genome. We further demonstrate that deletion of bfmS causes substantially reduced virulence in lettuce leaf, reduced cytotoxicity, enhanced invasion, and reduced bacterial survival during acute mouse lung infection. Intriguingly, specific missense mutations, which occur naturally in the bfmS gene in P. aeruginosa cystic fibrosis (CF) isolates such as DK2 strains and RP73 strain, can produce BfmS variants (BfmS_L181P, BfmS_L181P/E376Q, and BfmS_R393H) that no longer repress, but instead activate BfmR. As a result, BfmS variants, but not the wild-type BfmS, inhibit the rhl QS system. This study thus uncovers a previously unexplored signal transduction pathway, BfmS/BfmR/RhlR, for the regulation of rhl QS in P. aeruginosa. We propose that BfmRS TCS may have an important role in the regulation and evolution of P. aeruginosa virulence during chronic infection in CF lungs.     

Altered antibiotic transport in OmpC mutants isolated from a series of clinical strains of multi-drug resistant E. coli

Antibiotic-resistant bacteria, particularly gram negative species, present significant health care challenges. The permeation of antibiotics through the outer membrane is largely effected by the porin superfamily, changes in which contribute to antibiotic resistance. A series of antibiotic resistant E. coli isolates were obtained from a patient during serial treatment with various antibiotics. The sequence of OmpC changed at three positions during treatment giving rise to a total of four OmpC variants (denoted OmpC20, OmpC26, OmpC28 and OmpC33, in which OmpC20 was derived from the first clinical isolate). We demonstrate that expression of the OmpC K12 porin in the clinical isolates lowers the MIC, consistent with modified porin function contributing to drug resistance. By a range of assays we have established that the three mutations that occur between OmpC20 and OmpC33 modify transport of both small molecules and antibiotics across the outer membrane. This results in the modulation of resistance to antibiotics, particularly cefotaxime. Small ion unitary conductance measurements of the isolated porins do not show significant differences between isolates. Thus, resistance does not appear to arise from major changes in pore size. Crystal structures of all four OmpC clinical mutants and molecular dynamics simulations also show that the pore size is essentially unchanged. Molecular dynamics simulations suggest that perturbation of the transverse electrostatic field at the constriction zone reduces cefotaxime passage through the pore, consistent with laboratory and clinical data. This subtle modification of the transverse electric field is a very different source of resistance than occlusion of the pore or wholesale destruction of the transverse field and points to a new mechanism by which porins may modulate antibiotic passage through the outer membrane.     

Von Hippel-Lindau disease: identification of deletion mutations by pulsed-field gel electrophoresis

Von Hippel-Lindau disease (VHL) is an inherited multisystem neoplastic disorder. We prepared a 2.5-megabase (Mb) restriction map of the region surrounding the VHL gene and identified and characterized overlapping deletions in three unrelated patients affected with VHL. The smallest nested deletion (100 kb) was located within a 510-kb NruI fragment detected by 19–63. The rearrangements detected will be useful in isolating and evaluating candidate cDNAs for the VHL gene. The detailed physical map will be useful in studying the organization and structure of genes in the VHL region.     

黑曲霉（Aspergillus niger）和运动发酵单胞菌（Zymomonas mobilis）共固定化细胞酒精发酵的研究

本研究将生产上常用的糖化菌（Ａｓｐ．ｎｉｇｅｒ３．４３０９）与酒精产生菌（Ｚ．ｍｏｂｉｌｉｓ）共同固定在海藻酸钙凝胶中,以淀粉为原料进行固定化细胞酒精发酵实验,并就ｐＨ、温度、通气量。淀粉浓度等对发酵的影响进行了探讨,认为当ＰＨ为５．０～５．５,温度为３０℃～３３℃时乙醇产率最高（Ｙｐ／ｓ可达０．３９）。而淀粉浓度、通气量对共固定化细胞酒精发酵有显著影响,二者存在着较强的交互作用,在发酵过程中根据淀粉粘度情况调节通气量将有助于提高乙醇产率。     

截短的猪繁殖与呼吸综合征ORF5基因及重组蛋白的原核表达

通过PCR方法从重组质粒pGEM-ORF5扩增得到缺失N端疏水序列的基因片段dORF5(deleting ORF5)。将dORF5克隆至原核高效表达载体pGEX-4T-1,在E.coliRosetta细胞中成功表达了重组蛋白GST-dORF5。用Western blotting鉴定表达蛋白,证明dORF5基因得到表达。本试验得到的重组蛋白,为进一步研究PRRS病毒结构蛋白的结构和功能奠定了基础。