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Refolding, purification and characterization of the major cockroach Cr PI allergen from Escherichia coli inclusion bodies

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Refolding, purification and characterization of the major co...

华东六省一市生物化学与分子生物学会2003年学术交流会

作者： Gao Bo,Liu Zhigang ,Hong Shiyi, Luo Shiwen (College of Life Sciences, shenzhen University.shenzhen,518060 shenzhen key laboratory of microbial genetic engineering.shenzhen 518060

Airborne cockroach allergens play an important role in allergic diseases, especially asthma. A lambda ExcellcDNA library was constructed from Periplaneta Americana nymph mRNA. The gene encoding Cr PI, the major al-lergen of P. America nymph was isolated with sera from subjects allergic to cockroach and subcloned into the fusionexpression vector of pGEX-5X-1 (Pharmacia), resulting in a GST-Cr P1 fusion gene construction (pGEX-5X-1/CrP1). This plasmid was transformed into and expressed by the Escherichia coli strain BL21. Most of the expressedfusion protein was found in the insoluble fraction (inclusion body) of the cell lysate.. After intensive washing, the

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Construction and primary characterization of cDNA expression library of Artemisia apiacea pollen

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Construction and primary characterization of cDNA expression...

华东六省一市生物化学与分子生物学会2003年学术交流会

作者： Liu Zhigang, Gao Bo, He YiHua,Su Dongming, Zhong suping, Wu xiaoman College of Life Sciences, shenzhen University.shenzhen,518060 shenzhen key laboratory of microbial genetic engineering.shenzhen, 518060 Second Affiliated Hospital of Guangzhou Medical College, Guangzhou,510260)

Backgrouds:Allergic diseases including extrinsic asthma(allergic asthma),allergic rhinitis *** defined as a

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Novel multi-objective optimization algorithm

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系统工程与电子技术：英文版 2014年第4期25卷 697-710页

作者： Jie Zeng Wei Nie College of Information engineering Shenzhen University Modern Communication and Information Processing key laboratory Shenzhen University

Many multi-objective evolutionary algorithms(MOEAs)can converge to the Pareto optimal front and work well on two or three objectives, but they deteriorate when faced with manyobjective problems. Indicator-based MOEAs, which adopt various indicators to evaluate the fitness values(instead of the Paretodominance relation to select candidate solutions), have been regarded as promising schemes that yield more satisfactory results than well-known algorithms, such as non-dominated sorting genetic algorithm(NSGA-II) and strength Pareto evolutionary algorithm(SPEA2). However, they can suffer from having a slow convergence speed. This paper proposes a new indicatorbased multi-objective optimization algorithm, namely, the multiobjective shuffled frog leaping algorithm based on the ε indicator(ε-MOSFLA). This algorithm adopts a memetic meta-heuristic,namely, the SFLA, which is characterized by the powerful capability of global search and quick convergence as an evolutionary strategy and a simple and effective ε-indicator as a fitness assignment scheme to conduct the search procedure. Experimental results, in comparison with other representative indicator-based MOEAs and traditional Pareto-based MOEAs on several standard test problems with up to 50 objectives, show that ε-MOSFLA is the best algorithm for solving many-objective optimization problems in terms of the solution quality as well as the speed of convergence.

关键词： evolutionary algorithm multi-objective optimization shuffled frog leaping algorithm(SFLA) convergence

Expression, Characterization and Purification of the Major Cockroach Cr PI Allergen

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Expression, Characterization and Purification of the Major C...

中国蛋白质组学首届学术大会

作者： Gao Bo, Liu Zhigang,Hong Shiyi, Luo Shiwen (College of Life Sciences, shenzhen University,shenzhen 518060 shenzhen key laboratory of microbial genetic engineering, shenzhen 518060)

Backgroud: Airborne cockroach allergens play an important role in allergic diseases, especially *** and therapy of cockroach allergy would benefit from the availability of defined recombinant ***: The purpose of this study was to express and purify recombinant Cr PI, a major allergen of ***: A lambda Excell cDNA expression library was constructed from Periplaneta Americana nymph *** sera from subjects allergic to cockroach,the cDNA library was immunoscreened and the isolated Cr PIgene was then subcloned into the fusion expression vector of pGEX-5X- 1 (Pharmacia).By means of WesternBlotting, the IgE-binding ability of the recombinant allergen was evaluated. Most of the expressed fusion pro-tein was found in the insoluble fraction (inclusion body) of the cell lysate. After intensive washing, the proteins

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A PSTAIRE CDK-like protein localizes in nuclei and cytoplasm of Physarum polycephalum and functions in the mitosis

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Cell Research 2004年第2期14卷 169-175页

作者： Gui Ying LI~(1,2), Miao XING~(1,*), Bo HU~(1,3) ~1College of Life Science, shenzhen key laboratory of microbial Gene engineering, shenzhen University, shenzhen 518060, China. ~2Department of Immunology, School of Basic Medical Sciences, Jilin University, Changchun 130021, China ~3Institute of genetics and Cytology, Northeast Normal University, Changchun 130024, China. College of Life Science Shenzhen Key Laboratory of Microbial Gene Engineering Shenzhen University Shenzhen 518060 China.

CDKs play key roles in controlling cell cycle progression in all eukaryotes. In plants, multiple CDKs are present, among which the best characterized CDKs are PSTAIRE CDKs. In this study, we carried out Western blot, immunoelectron microscopy and antibody treatment with an anti-PSTAIRE monoclonal antibody to explore the subcellular localization and functions of PSTAIRE CDKs in Physarum polycephalum. The results of Western blot and immunoelectron microscopy showed that in P. polycephalum, a PSTAIRE CDK-like protein was 34 kD in molecular weight and located in both nuclei and cytoplasm. In nuclei, the protein was mainly associated with chromosomes and nucleoli. The expression of the PSTAIRE CDK-like protein in both the plasmodia and nuclei showed little fluctuation through the whole cell cycle. When treated with an anti-PSTAIRE monoclonal antibody at early S phase, the cells were arrested in S phase, and the mitotic onset of P. polycephalum was blocked for about 1 h when treated at early G2 phase. Our dat

关键词： PSTAIRE CDK-like protein Physarum polycephalum mitosis immunoelectron microscopy.

Regulation of eight avr genes by hrpG and hrpX in Xanthomonas campestris pv. campestris and their role in pathogenicity

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Progress in Natural Science:Materials International 2006年第12期16卷 1288-1294页

作者： XU Rongqi~ 1,3 , LI Xianzhen~3, WEI Hongyu~3, JIANG Bole~2, LI Kai~3, HE Yongqiang~ 2,3 , FENG Jiaxun~ 1,3 and TANG Jiliang~ 1,2 **(1. Guangxi key laboratory of Subtropical Bioresources Conservation and Utilization 2. The key laboratory of Ministry of Education for microbial and Plant genetic engineering 3. College of Life Science and Technology, Guangxi University, Nanning 530004, China) Guangxi key laboratory of Subtropical Bioresources Conservation and Utilization Guangxi University Nanning 530004 China. College of Life Science and Technology Guangxi University Nanning 530004 China. key laboratory of Ministry of Education for microbial and Plant genetic engineering Guangxi University Nanning 530004 China

Eight putative avirulence genes in Xanthomonas campestris pv. campestris (Xcc) strain 8004 were characterized by Tn5gusA5 mutagenesis and gene expression analysis. The virulence test of mutants on Chinese radish showed that all mutants in individual avr genes except avrBs2 mutant were not significantly different from the wild type in virulence. The avrBs2 mutant showed reduced virulence and bacterial growth in planta. Gene expression analysis using β-glucuronidase as reporter indicated that avrBs1.1,avrBs1,avrXccB,avrXccC,avrXccE1 were regulated by hrpG, whereas avrXccA1, avrXccA2 and avrBs2 were not. RT-PCR analysis showed that all hrpG-regulated genes except avrBs1 were also regulated by hrpX. In addition, it was demonstrated that avrBs1 was responsible for elicitation of a type III dependent hypersensitive reaction (HR) on nonhost plant pepper ECW-10R, and wild type Xcc 8004 was unable to cause HR on pepper ECW-20R.

关键词： Xanthomonas campestris pv. campestris avirulence gene virulence expression analysis.

Genomic research for important pathogenic bacteria in China

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Science China(Life Sciences) 2009年第1期52卷 50-63页

作者： YANG RuiFu1,GUO XiaoKui2,YANG Jian3,JIANG YongQiang1,PANG Bo4,CHEN Chen5,YAO YuFeng2,QIN JinHong2 & LI QingTian2 1 laboratory of Analytical Microbiology,State key laboratory of Pathogen and Biosecurity,Beijing Institute of Microbiology and Epidemiology,Beijing 100071,China 2 Department of Medical Microbiology and Parasitology,Institutes of Medical Sciences,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China 3 State key laboratory for Molecular Virology and genetic engineering,Institute of Pathogen Biology,Chinese Academy of Medical Sciences,Beijing 100176,China 4 State key laboratory for Infectious Disease Prevention and Control,Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China 5 Beijing Genomics Institute at shenzhen,shenzhen 518083,China laboratory of Analytical Microbiology State Key Laboratory of Pathogen and Biosecurity Beijing Institute of Microbiology and Epidemiology Beijing 100071 Chin

Rapid accumulation of bacterial genomic data offered an unprecedented opportunity to understand bacterial biology from a holistic view of *** can thus closely look at the way in which a pathogen is evolved,and these data has been applied to molecular epidemiology and microbial forensics,and screening of novel diagnostic,vaccine and drug *** newly developed high-throughput low-cost sequencing technologies,such as 454,Solexa and SOLiD,will promote the acquisition and application of genomic data in new research areas that we dared not imagine previously,such as the metagenomics of human gastric-intestinal tract,for better and comprehensive understanding of human health and disease.

关键词： genome pathogenic bacteria sequencing application

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Dissecting and Exploiting Nonribosomal Peptide Synthetases

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Acta Biochimica et Biophysica Sinica 2004年第4期36卷 243-249页

作者： Qing-Tao SHEN, Xiu-Lan CHEN, Cai-Yun SUN, and Yu-Zhong ZHANG*State key laboratory of microbial Technology, Shandong University, Ji’nan 250100, China State key laboratory of microbial Technology Shandong UniversityJi'nan 250100 China

A large number of therapeutically useful cyclic and linear peptides of bacteria or fungal origin are synthesized via a template-directed, nucleic-acid-independent nonribosomal mechanism. This process is carried out by mega-enzymes called nonribosomal peptide synthetases (NRPSs). NRPSs contain repeated coordinated groups of active sites called modules, and each module is composed of several domains with different catalytic activities. The familiarity to these domains lays base for the future genetic engineering of NRPSs to generate entirely “unnature” products. The details about NRPSs domain structures and the exploi- tation of NRPSs are described in this review.

关键词： nonribosomal peptide synthetase domain combinatorial biosynthesis

Cloning and expression of Haematococcus pluvialis fatty acid synthsis genes and its relationship with fatty acid accumulation

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Cloning and expression of Haematococcus pluvialis fatty acid...

中国藻类学会第八次会员大会暨第十六次学术讨论会

作者： An-ping Lei~1,Huan Chen~1,Chao-gang Wang~1,Zhang-li Hu and Jiang-xin Wang~(2,*) 1 shenzhen key laboratory for Marine Bio-resource and Eco-environment,College of Life Science,shenzhen University,shenzhen 518060,P.R.China 2 Biodesign Institute,Arizona State University,Tempe,AZ 85287,USA

＜正＞Biofuel has been an incredibly hot research topic recently in both academia and *** feedstocks include crops like corn,soybeans,flaxseed rapeseed/canola,and microorganisms,such as algae,yeast,bacteria,and *** them,microalgae become more attractive due to their rapid growth,high oil content, high density culture and with no competition with food resources and agricultural land. To blast through the bottleneck of high biofuel production cost from microalgae, synthesis biology techniques are highly required to improve content of algal fatty acids in *** fatty acid biosynthesis pathway genes have been all cloned and biosynthesis pathway was built up in some higher plants and algae,the molecular mechanism for its regulation is far away from *** this end,we cloned main key genes for fatty acid biosynthesis in Haematococcus pluvialis,a green alga with high commercial values,and investigated relationships between their expression alternation and fatty acid profiling detected by GOMS under different stress treatments, *** depletion,salinity,high or low *** results showed that high temperature,high salinity,and nitrogen depletion treatments played significant role in promoting algal fatty acid *** analysis showed that acyl carrier protein（ACP）,3-ketoacyl-ACP-synthase（KAS）,and acyl-ACP thioesterase（FATA） gene expression had significant or highly significant correlations with monounsaturated fatty acid synthesis（positive） and polyunsaturated fatty acid synthesis（negative）. We proposed that ACP,KAS and FATA in Haematococcus play an important role in fatty acid synthesis and may be rate limiting genes,which could be modified for the further study of genetic engineering,metabolic engineering to improve algae biofuel quality and production.

关键词： Haematococcus pluvialis fatty acid synthesis fatty acid synthase gene gene expression