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Identification and Characterization of hmr19 Gene Encoding a Multidrug Resistance Efflux Protein from Streptomyces hygroscopicus *** Strain 10-22

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 519-528页

作者： Lei QIN , Heng-An WANG1 , Zhong-Qin WU, Xiao-Feng ZHANG, Mei-Lei JIN,Zi-Xin DENG2, and Guo-Ping ZHAOshanghaiResearch Center of Biotechnology, shanghai Institute for Biological Sciences, Chinese Academy of Sciences,Graduate school of the Chinese Academy of Sciences, shanghai 200233, china 1school of agriculture and biology, shanghai jiaotong university, shanghai 201101, china 2school of Life Science and Biotechnology, shanghai jiaotong university, shanghai 200030, china shanghaiResearch Center of Biotechnology Shanghai Institute for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences school of agriculture and biology Shanghai Jiaotong University

The hmr19 gene was cloned from Streptomyces hygroscopicus subsp. yingchengensis strain 10- 22, a bacterium strain producing agricultural antibiotics. Sequence similarity comparison indicates that hmr19 gene may encode a predicted protein with 14 putative transmembrane α-helical spanners, belonging to the drug:H+ antiporter-2 family of the major facilitator superfamily. The expression of hmr19 in the mycelium of strain 10-22 was detected by Western blotting analysis. Gene replacement technology was employed to construct an hmr19 disruption mutant. The growth inhibition test against different antibiotics indicated that the mutant strain was 5–20 fold more susceptible to tetracycline, vancomycin and mitomycin C than the parental wild type strain. The mutant took up tetracycline much faster and accumulated more antibiotics than the wild type strain 10-22. While with the addition of an energy uncoupler, carbonyl cyanide m- chlorophenylhydrazone, the characteristics of the accumulation of [3H]tetracycline in these two strains were almost the same. It was thus concluded that hmr19 encoded a multidrug resistance efflux protein.

关键词： multidrug resistance (MDR) gene replacement (knockout) Streptomyces Streptomyces hygroscopicus subsp. yingchengensis strain 10-22

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Construction of a genetic map with SRAP markers and localization of the gene responsible for the first-flower-node trait in cucumber (Cucumis sativus L.)

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Progress in Natural Science:Materials International 2005年第5期15卷 407-413页

作者： PAN Junsong*, WANG Gang*, LI Xiaozun, HE Huanle, WU Aizhong and CAI Run**(agriculture and biology school, shanghai jiaotong university, shanghai 201101, china) agriculture and biology school Shanghai Jiaotong University Shanghai 201101 Chine

With an F 2 population from the cross of two cucumber inbred lines, S06 and S52, sequence-related amplified polymorphism (SRAP) was used to construct a genetic linkage map in cucumber (Cucumis sativus L.). Sixty-four SRAP primer combinations generated 108 polymorphic bands in the F 2 population analysis. The average of polymorphic bands produced by one primer pair was 1.5, and the maximum was 5. Using Mapmaker 3.0, a linkage map was constructed, which consisted of 77 SRAP markers distributed in nine linkage groups (LOD≥3.0) and spanned 1114.2 cM with an average interval of 14.5 cM between markers. The gene for the first-flower-node trait, termed ffn, was mapped to linkage group Ⅸ, flanked by DC1EM5 and ME7EM2A at 10.3 cM and 12.1 cM distance, respectively.

关键词： cucumber (Cucumis sativus L.) SRAP (sequence-related amplified polymorphism) linkage map first flower node.

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Localization of genes for lateral branch and female sex expression and construction of a molecular linkage map in cucumber (Cucumis sativus L.) with RAPD markers

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Progress in Natural Science:Materials International 2005年第2期15卷 48-53页

作者： LI Xiaozun1*, PAN Junsong1*, WANG Gang1, TIAN Libo3, SI Longting3, WU Aizhong2** and CAI Run1**(1. agriculture and biology school, shanghai jiaotong university, shanghai 201101, china 2. shanghai Academy of Agricultural Science, shanghai 201106, china 3. Horticulture school, Shenyang Agricultural university, Shenyang 110161, china) agriculture and biology school Shanghai Jiaotong University Shanghai 201101 China. Horticulture school Shenyang Agricultural University Shenyang 110161 China. shanghai Academy of Agricultural Science Shanghai 201106 China

A cucumber (Cucumis sativus L.) molecular linkage map, including 79 random-amplified polymorphic DNAs (RAPD) and two genes , lb for lateral branch and f for female sex expression, is constructed from a cross between a line, S52, with weak lateral growing ability and staminate from Dabieshan Mountains area in china and another line, S06, with strong lateral growing ability and gynoecious from Europe. The map contains nine linkage groups and spans 1110.0 cM with an average distance of 13.7 cM between loci. The lb locus is located in a longer linkage group LG-2 and flanked by two markers, OP-Q5-1 and OP-M-2-2, at 9.3 cM and 15.9 cM, respectively. In the meantime, the RAPD loci, OP-Q5-2 and BC151, in a short linkage group were found to flank f at 13.7 cM and 13.4 cM, respectively. The construction of RAPD map has paved a way for further study of the genes for lateral branch, female sex expression and other agronomic traits in cucumber.

关键词： cucumber (Cucumis sativus L.) lateral branch gynoecy RAPD genetic mapping

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Calculation of the molecular exchanging energy of binary surfactants system on the surface monolayer of aqueous solution

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Science china Chemistry 2007年第4期50卷 468-475页

作者： WANG ZhengWu1 & YI XiZhang2 1 Department of Food Sciences & Technology, school of agriculture and biology, shanghai Jiao Tong university, shanghai 200240, china 2 Institute of Theoretical Chemistry, Shandong university, Jinan 250100, china 1. Department of Food Sciences & Technology School of Agriculture and Biology Shanghai Jiao Tong University Shanghai 200240 China 2. Institute of Theoretical Chemistry Shandong University Jinan 250100 China

By using the binary anionic/cationic surfactants system CH3(CH2)nOSO_3/CH3(CH2)nN+(CH3)3 as an ex-ample, the molecular exchanging energy (ε) of adsorption on the surface monolayer of aqueous solu-tion has been studied. ε can be obtained with two methods. One is from the relationship between ε and the molecule interaction parameter (β). This relationship is founded by considering that the adsorption of mixed surfactants on the surface monolayer of solution satisfies the dimensional crystal model condition under which β can be obtained by testing the surface tension of solution. The other is directly from the molecular structure of surfactants with the Lennard-Jones formula. The results for the studied system show that these two methods coincide well.

关键词： surfactant molecule exchanging energy dimensional crystal model

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Comparative proteomic analysis of the response in resistant and susceptible maize inbred lines to infection by Curvularia lunata

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Progress in Natural Science:Materials International 2009年第7期19卷 845-850页

作者： Xiuli Huang 1, Lixing Liu 1, Jie Chen Yuhong Zhai Department of Plant Science, school of agriculture and biology, Key Laboratory of Microbial Metabolism, Ministry of Education, shanghai jiaotong university, shanghai 200240, china [a]Department of Plant Science School of Agriculture and Biology Key Laboratory of Microbial Metabolism Ministry of Education Shanghai Jiaotong University Shanghai 200240 China

Proteins differentially expressed from maize leaves in response to the infection by Curvularia lunata strain CX-3 were identified through a high-resolution two-dimensional gel electrophoresis (2-DE) method. Two inbred lines, 78599-1 and E28, were used, respectively, as resistant and susceptible lines to CX-3 infection. Proteins were extracted from the fourth leaves of sixor seven-leaf stage plants sampled at 24, 36, 48, 60, and 72 h after inoculation with CX-3. Twenty-seven differentially expressed protein spots resolved on the 2-DE gels were identified by MALDI-TOF MS/MS. The results showed that these proteins are associated with photosynthesis, respiration, oxidative and drought stress tolerance as well as signal transduction in maize. Among stress-related proteins, the 22 kDa drought-inducible protein, putative glutathione peroxidase (GPX), and translation initiation factor (eIF-5A) were up-regulated in the resistant inbred line and were implicated in host defense response to C. lunata infection. It suggests that drought-inducible and oxidation stress-related proteins might directly contribute to maize resistance to C. lunata.

关键词： Curvularia lunata Maize Proteomics Two-dimensional electrophoresis (2-DE)

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Construction of a BAC library from cucumber (Cucumis sativus L.) and identification of linkage group specific clones

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Progress in Natural Science:Materials International 2008年第2期18卷 143-147页

作者： Yuan Guan a,1,Qi Chen b,1,Junsong Pan a,Zheng Li a,Huanle He a,Aizhong Wu a,Rentao Song b,Run Cai a,a school of agriculture and biology,shanghai Jiao Tong university,shanghai 200240,china b school of Life Sciences,shanghai university,shanghai 200444,china school of agriculture and biology Shanghai Jiao Tong University Shanghai 200240 China school of Life Sciences Shanghai University Shanghai 200444 China

A bacterial artificial chromosome (BAC) library consisting of 19,200 clones with an average insert size of 105 kb has been constructed from a cucumber (Cucumis sativus L.) inbred line S94,derived from a cultivar in North china. The entire library was equivalent to approximately 5 haploid cucumber genomes. To facilitate chromosome engineering and anchor the cucumber genetic linkage map to its chromosomes,15 sequence-characterized ampli?ed regions (SCAR) and seven simple sequence repeats (SSR) markers from each link-age group of cucumber were used to screen an ordered array of pooled BAC DNA with polymerase chain reaction (PCR). Fifteen mark-ers gave at least two positive clones. As a result,22 BAC clones representing 7 linkage groups of cucumber were identified,which further validated the genome coverage and utility of the library. This BAC library and linkage group specific clones provide essential resources for future research of the cucumber genome.

关键词： Cucumber BAC library Molecular markers PCR screening Specific clones

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QTL molecular marker location of powdery mildew resistance in cucumber (Cucumis sativus L.)

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Science china(Life Sciences) 2008年第11期51卷 1003-1008页

作者： LIU LongZhou1,2, CAI Run1, YUAN XiaoJun1, HE HuanLe1 & PAN JunSong1 1 school of agriculture and biology, shanghai jiaotong university, shanghai 200240, china 2 Horticultural Research Institute, shanghai Academy of Agricultural Sciences( (SAAS)), shanghai Key Lab of Protected Horti-cultural Technology, shanghai, 201106, china 1. school of agriculture and biology Shanghai Jiaotong University Shanghai 200240 China 2. Horticultural Research Institute Shanghai Academy of Agricultural Sciences (SAAS) Shanghai Key Lab of Protected Horticultural Technology Shanghai 201106 China

The cucumber lines, S94 (Northern china open-field type, powdery mildew (PM) susceptible) and S06 (European greenhouse type, PM resistant), and their F6:7 populations were used to investigate PM re-sistance under seedling spray inoculation in 2005/Autumn and 2006/Spring. QTL analysis was under-taken based on a constructed molecular linkage map of the corresponding F6 population using com-posite interval mapping. A total of four QTLs (pm1.1, pm2.1, pm4.1 and pm6.1) for PM resistance were identified and located on LG 1, 2, 4 and 6, respectively, explaining 5.2%-21.0% of the phenotypic variation. Three consistent QTLs (pm1.1, pm2.1 and pm4.1) were detected under the two test conditions. The QTL pm6.1 was only identified in 2005/Autumn. The total phenotypic variation explained by the QTLs was 52.0% and 42.0% in 2005/Autumn and 2006/Spring, respectively. Anchor markers tightly linked to those loci (<5 cM) could lay a basis for both molecular marker-assisted breeding and map-based gene cloning of the PM-resistance gene in cucumber.

关键词： cucumber powdery mildew QTL F6:7families seedling

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Identification and Characterization of a Rat Novel Gene RSEP4 Expressed Specifically in Central Nervous System

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Acta Biochimica et Biophysica Sinica 2004年第7期36卷 501-507页

作者： Xi-Dao WANG1,2, Ling-Wei KONG1, Zhi-Qin XIE1, Yu-Qiu ZHANG3, Zhi-Xin LIN2, Zhi-Qi ZHAO3,Lei YU4, and Nai-He JING1* 1 Laboratory of Molecular Cell biology, Laboratory of Stem Cell biology, Institute of Biochemistry and Cell biology, shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Graduate school of the Chinese Academy of Sciences, shanghai 200031, china school of Life Science and Biotechnology, 2 shanghai jiaotong university, shanghai 200030, china Institute of Neurobiology, Fudan university, shanghai 200433, china 3 4 Department of Cell biology, Neurobiology and Anatomy, College of Medicine, university of Cincinnati, Cincinnati, Ohio, USA Laboratory of Molecular Cell biology and Stem Cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences Shanghai China

The low-abundantly expressed genes composed the majorities of the mRNAs expressed in the central nervous system (CNS), and were thought to be important for the normal brain functions. Through differential screening a low-abundance cDNA sublibrary with mRNA from neuropathic pain of chronic constriction injury (CCI) model, we have identified a novel rat gene, rat spinal-cord expression protein 4 gene (RSEP4). The total length of RSEP4 cDNA is 2006 bp, with a 501 nucleotide open reading frame (ORF) that encodes a 167 amino acid polypeptide. Northern blot revealed that RSEP4 was expressed specifically in the CNS. In situ hybridization showed that the mRNA of RSEP4 was strongly expressed in the CA1, CA2, CA3 and DG regions of hippocampus, the Purkinje cells of cerebellum, and the small sensory neurons of dorsal horn and large motor neurons of ventral horn of spinal cord. Over-expression of RSEP4-EGFP fusion protein in the human embryonic kidney 293T cells showed that RSEP4 protein was mainly localized in the cell cytoplasm. These results suggest that RSEP4 may play some roles in the CNS.

关键词： central nervous system (CNS) rat Northern blot in situ hybridization

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The cell cycle related apoptotic susceptibility to arsenic trioxide is associated with the level of reactive oxygen species

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Cell Research 2004年第1期14卷 81-85页

作者： Fei GAO~1, Jing YI~1, Jing Qi YUAN~2, Gui Ying SHI~1, Xue Ming TANG~(1,*) ~1Department of Cell biology, shanghai Second Medical university, shanghai 200025, china ~2Department of Automation, shanghai jiaotong university, shanghai 200031, china. Tang XM Shanghai Med Univ 2 Dept Cell Biol Shanghai 200025 Peoples R China

Double staining flow cytometry was performed using 7-amino actinomycin D and 6-carboxy-2’,7’-dichlorodihydrofluorescein diacetate, to detect the level fluctuation of reactive oxygen species （ROS） during the cell cycle of normal NB4 cells. Our results showed that NB4 cells possessed higher level of ROS in G2/M phase than in G1 and S phases. Double staining flow cytometry, with TdT mediated dUTP nick end labeling （Tunel） and propidium iodide （PI）, indicated that As2O3 （2μM） could induce apoptosis in NB4 cells prevailingly from G2/M phase, and this efficacy was enhanced upon co-administration of 2, 3-dimethoxy-1, 4-naphthoquinone （DMNQ）（2.5μM） which could produce the endogenous ROS. These results suggested that different ROS level in different cell cycle phases of NB4 cells might determin the selective induction of G2/M apoptosis and the cells’ susceptibility to apoptosis by As2O3.

关键词： arsenic trioxide apoptosis cell cycle reactive oxygen species (ROS).

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Identification of Soybean Resources of Resistance to Aphids

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Agricultural Sciences in china 2009年第8期8卷 979-984页

作者： WU Tian-long,MA Xiao-hong,YAO Lu-ming and WANG Biao school of agriculture and biology,shanghai Jiao Tong university,shanghai 200240,P.R.china [a]school of agriculture and biology Shanghai Jiao Tong University Shanghai 200240 P. R. China

Four soybean [Glycine max （L.） Merr.] cultivars with soybean aphid resistance （Aphis glycines Matsmura）, p189, P203, P574, and P746, were identified in field test, choice test, and non-choice test, The grade of resistance to aphids and the damage index of P189, P203, and P746 were significantly different from the susceptible cultivars （P=0.05）. P574 and P746 showed antibiosis resistance, preventing aphids from reproducing on the plants. P203 showed antixenosis resistance, preventing aphids from reproducing in field test and choice test, but susceptible in non-choice test. Population development on plants was significantly different in field test, choice test, and non-choice test, which was caused by different selective pressures.

关键词： Aphis glycines Matsmura resistance to aphids antibiosis antixenosis

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