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CasHRAC:Cas9-facilitated homologous recombination for efficient assembly of circular DNAs up to several hundred kilobases

CasHRAC:Cas9-facilitated homologous recombination for effici...

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中国生物工程学会第六次全国会员代表大会暨第九届学术年会

作者：周见庭 Ronghai Wu Xiaoli Xue Zhongjun Qin 上海植物生理生态研究所合成生物学重点实验室 Key Laboratory of Synthetic Biology Institute of Plant Physiology and EcologyShanghai Institutes for Biological SciencesChinese Academy of Sciences

Standardized DNA assembly have enabled a broad range of synthetic biology ***,the assembly of large constructs up to several hundred kilobases has been challenging,mainly due to the complex in vitro manipulation of long DNA ***,we describe a new method designated "CasHRAC(Cas9-facilitated homologous recombination for assembly of circular DNAs)",which allows direct usage of multiple large plasmids(> 100 kbp) for one-step DNA assembly in *** large plasmids are gathered into the same yeast cell by protoplast fusion,and be cut by the RNA-guided Cas9 nuclease to release the linear DNA segments to join in assembly by taking advantage of the native homologous recombination *** a proof-of-concept,we apply CasHRAC for assembly of plasmids with various length of insertions(10～298 kbp) into four DNA constructs,and the largest assembly is 660 kbp and contains all published Escherichia coli essential *** method complements and improves the current methods for large DNA assembly,especially in the whole genome assembly.

关键词： DNA CasHRAC:Cas9-facilitated homologous recombination for efficient assembly of circular DNAs up to several hundred kilobases

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QCFVA:A tool for qualitative classification of reactions based on flux variability analysis

QCFVA:A tool for qualitative classification of reactions bas...

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中国生物工程学会第六次全国会员代表大会暨第九届学术年会

作者：李培顺 Hongwu Ma Teng Huang 中国科学院天津工业生物技术研究所 Tianjin Institute of Industrial Biotechnology Chinese Academy of Sciences

Background:Flux variability analysis is usually used to determine the minimum and maximum flux values that the reactions in the metabolic network can carry under various simulation *** several approaches have been proposed for that purpose,they are computationally too inaccurate to qualitatively classify reactions in genome-scale metabolic network models(GSMs).Therefore,developing preciser methods to analysize robustness of metabolic network models is ***:We develop a new tool for qualitative classification of reactions base on flux variability analysis called QCFVA,which overcomes the problem of calculation accuracy of the exiting *** implementation of QCFVA was mainly divided into three steps:Firstly,the target reaction,such as biomass growth,was set as the objective function and FBA was used to maximize the flux of the target reaction(mentioned as the first simulation);Secondly,the flux of target reaction was set to the maximum value calculated in the first simulation;Finally,all the n reactions were considered as objective function in turn and FBA was again used to calculate the maximum and minimum fluxes of each reaction in GSMs,***:We show that the improved implementation of QCFVA makes classification of reactions,calculation of the bounds on the reaction fluxes and flux balance impact degree(FBID) of single reaction deletion more exactly,greatly expanding the application of flux variability analysis in systems biology from prediction of genetic manipulation targets in genome scale to integration of high-throughput biological data to construct multi-scale whole-cell network model.

关键词： flux variability analysis genome-scale metabolic network reaction classification

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Hypoglycemic Effect of Black Garlic in Experimental Diabetic Rats

Hypoglycemic Effect of Black Garlic in Experimental Diabetic...

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中国生物工程学会第六次全国会员代表大会暨第九届学术年会

作者： LIU Yu-feng GUO Chun-jing SUN Yu-feng 杨庆丽 HAN Qiu-jv Daqing Branch of Heilongjiang Academy of Sciences Heilongjiang Academy of Sciences 黑龙江省科学院大庆分院生物新技术研究所 Heilongjiang Land Reclamation Vocational College

Objective To investigate the effect and mechanism of the "black garlic"(Black garlic) on the body weight and the blood glucose in diabetic rats induced by *** The diabetic rat model was established by intraperitoneal injection of 50 mg/kg *** garlic was administrated to the rats by oral gavage for 30 *** weight and blood glucose level before and after 30-day oral gavage were determined *** Black garlic at the dose of 10 g/kg has no effect on body weight and blood glucose level of the control *** the high(10 g/kg) and the middle(5 g/kg) doses,body weight of the black garlic treated group declined,while the blood glucose level increased continuously compared with the untreated *** there were no significant differences between the two *** the low dose(2.5 g/kg),weight loss of the treated group was relieved and the blood glucose increased slowly compared with those of the untreated *** the differences were statistical *** This study indicates that proper consumption of black garlic reduces blood glucose level,and delays the development of *** studies are expected to explicate the mechanism of the black garlic.

关键词： Black garlic streptozotocin blood glucose

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The Effects of Cerium on Growth、Chlorophyll a Concent and Chlorophyll Fluorescence of Scendesmus obliquus

The Effects of Cerium on Growth、Chlorophyll a Concent and C...

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中国生物工程学会第六次全国会员代表大会暨第九届学术年会

作者：成杰 Ji Xiang Zhang Kaikai Zhao Xinyu Li Hongxia Gong Donghui 蔡禄内蒙古科技大学数理与生物工程学院 School of Mathematics Physicsand Biological EngineeringInner Mongolia University of Science and Technology Institute of Bioengineering and Technology Inner Mongolia Key Laboratory of Biomass-Energy Conversion

To investigate the effect of cerium（Ce） on Scendesmus obliquus,the growth,chlorophyll a content and chlorophyll fluorescence were studied by lab cultured *** from the present investigation noted that 0.50-5.00 mg/L Ce resulted in a considerable stimulation to the growth and the chlorophyll a content of Scendesmus obliquus,but concentrations above 50.0 mg/L appeared an inhibitory *** the growth and the chlorophyll a content were also at a maximum at a concentration of 5 mg/*** the donor and acceptor side of PSII reaction centers are sensitive to external *** our study,the OEC of PSII in the low concentration of Ce was not sensitive to Cerium,but sensitivity was intensely increased when treated with high concentration.Ψ_o and φ were significantly lower in the treatment than in the *** led to decrease in rate of electron transport（ET0/ABS and ET/TR）.An increase of VJ suggests that Ce inhibits electron transfer from Q to Q and subsequently results in accumulation of *** results provide not only new insights into cerium stress responses in Scendesmus obliquus but also a good foundation for realizing the cytological mechanism for the toxic effects of Ce on Scendesmus obliquus.

关键词： Scendesmus obliquus cerium growth chlorophyll a concent chlorophyll fluorescence

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Enzymatic synthesis of isoquercitrin with naringinase under microwave irradiation

Enzymatic synthesis of isoquercitrin with naringinase under ...

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中国生物工程学会第六次全国会员代表大会暨第九届学术年会

作者：朱丹 Yi-yuan Mei Xiao-hui Xu Chuan-qi Jin Jin-xian Wu An Gong Jun Wang 江苏科技大学生物技术学院 School of Biotechnology Jiangsu University of Science and Technology Sericultural Research Institute Chinese Academy of Agricultural Sciences

Isoquercitrin,a key synthetic intermediate,has a wide range of biological activities,for the production of enzymatically modified isoquercitrin（EMIQ） which has received FDA safety ***,the synthesis of isoquercitrin is still problematic,requires a long reaction time,is unstable and easily oxidizes *** order to establish an efficient bioprocess for obtaining isoquercitrin,a biosynthesis of isoquercitrin using the naringinase-catalyzed hydrothermal synthesis of rutin under microwave irradiation was first carried *** has been proved the feasibility of the free enzyme under microwave *** irradiation can effectively strengthen the enzymatic reaction,reduce the activation *** also can change the enzyme microscopic configuration and the reaction *** catalytic effect has a great thrust on the promotion of the enzymatic reactions in industrial *** micro-technology may be applied to the effective synthesis of other precious ***,a bright future and a better effect would be achieved by unremitting *** this study,enzymatic reactions were performed in a microwave reactor for isoquercitrin *** volume ratio of enzyme solution,ionic liquid and rutin dissolved by buffer solution in the enzymatic synthesis of isoquercitrin reaction system was 180:720:*** reaction was initiated by adding naringinase and the mixtures at various substrate concentrations,enzyme concentrations,reaction time,microwave power and types of ionic *** microwave irradiation,the mixture sampled for every 10 min from the 30 min to the 70 *** crude hydrolysis products of rutin were determined and verificated by HPLC and *** yield was calculated as follows Eq.（1）.Isoquercitrin yield（%）=moles of isoquercitrin/initial moles of rutin×100% （1）Reaction medium directly affects the reaction rate and the enzyme ***,ionic liquid has the strong ability of absorbing microwave,a

关键词： Microwave irradiation Naringinase Isoquercitrin Enzymatic synthesis

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Comparative Profiling of Coral Symbiont Microbial Communities from the South China Sea and Moorea

Comparative Profiling of Coral Symbiont Microbial Communitie...

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中国生物工程学会第六次全国会员代表大会暨第九届学术年会

作者： Maozhen Han Yanhai Gong Jiahuan Chen Kang Ning Bioinformatics Group of Single Cell Center Shandong Key Laboratory of Energy Genetics and CAS Key Laboratory of BiofuelsQingdao Institute of Bioenergy and Bioprocess TechnologyChinese Academy of SciencesQingdaoShandongChina 266101 College of Marine Life Ocean University of ChinaQingdaoShandongChina 266000 Bioinformatics Group of Single Cell Center Shandong Key Laboratory of Energy Genetics and CAS Key Laboratory of BiofuelsQingdao Institute of Bioenergy and Bioprocess TechnologyChinese Academy of SciencesQingdaoShandongChina 266101 Bioinformatics Group of Single Cell CenterShandong Key Laboratory of Energy Genetics and CAS Key Laboratory of BiofuelsQingdao Institute of Bioenergy and Bioprocess TechnologyChinese Academy of SciencesQingdaoShandongChina 266101 School of Life Science and TechnologyHuazhong University of Science and TechnologyWuhanHubeiChina 430070 School of Life Science and Technology Huazhong University of Science and TechnologyWuhanHubeiChina 430070

Coral reefs represent one of the richest ocean micro-scale environments that include diverse collection of both basic and advanced *** of the microbial communities that live with coral reefs could help to answer key ecological questions about coral *** connection of community structure and the environmental factors would also provide clues for important chemical *** this work,we have collected sequencing data associated with coral reef microbial communities collected from two distinct geological locations: the South China Sea and Moorea.A number of 25 coral reefs samples were collected,and categorized into 7 groups: Porites Iutea(Pl),Galaxea fascicularis(Gf),Acropora millepora(Am),Paopao Bay,Fore reef,Back reef and Fringing reef,from the datasets downloaded from NCBI(SRR82519 and SRP007926).The number of reads ranged from 554 to 23,858 in each ***,199,860 reads were remained after quality filtering and clustered into 2,149 different *** sequences were grouped under 31 high-level taxonomic groups at the phylum ***,the microbial communities from these two locations were largely different and the species with different abundances,especially cyanobacteria,were found to be indicators that could reflect human activities and coral reefs' health *** dominant phyla(Proteobacteria,Firmicutes and Bacteroidetes)comprised 58.6%and 71.0%of the Pl and Gf communities,***,for Am,three dominant phyla(Proteobacteria,Firmicutes and Deinococcus-Thermus)comprised 97.3%of the communities and 2 predominant phylum(Proteobacteria and Cyanobacteria)comprised 80.95%of the A,B,C,D,E and H sites ***,the results of NMDS,PCoA and Clustering analysis showed that these samples could be divided into three groups: Am group,Moorea group and Pl+Gf group,and the results were consistent with the result of ***,based on Kruskal-Wallis test among classes and the pairwise Wilcoxon test among

关键词： coral reefs microbial community symbiont cyanobacteria wellness indicators

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Comparative Profiling of Coral Symbiont Microbial Communities from the South China Sea and Moorea

Comparative Profiling of Coral Symbiont Microbial Communitie...

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中国生物工程学会第六次全国会员代表大会暨第九届学术年会

作者：韩毛振 Yanhai Gong Jiahuan Chen 宁康华中科技大学生命科学与技术学院 Bioinformatics Group of Single Cell Center Shandong Key Laboratory of Energy Genetics and CAS Key Laboratory of BiofuelsQingdao Institute of Bioenergy and Bioprocess TechnologyChinese Academy of Sciences School of Life Science and Technology Huazhong University of Science and Technology College of Marine Life Ocean University of China 华中科技大学生物信息系

Coral reefs represent one of the richest ocean micro-scale environments that include diverse collection of both basic and advanced *** of the microbial communities that live with coral reefs could help to answer key ecological questions about coral *** connection of community structure and the environmental factors would also provide clues for important chemical *** this work,we have collected sequencing data associated with coral reef microbial communities collected from two distinct geological locations:the South China Sea and Moorea.A number of 25 coral reefs samples were collected,and categorized into 7 groups:Porites Iutea(Pl),Galaxea fascicularis(Gf),Acropora millepora(Am),Paopao Bay,Fore reef,Back reef and Fringing reef,from the datasets downloaded from NCBI(SRR82519 and SRP007926).The number of reads ranged from 554 to 23,858 in each ***,199,860 reads were remained after quality filtering and clustered into 2,149 different *** sequences were grouped under 31 high-level taxonomic groups at the phylum ***,the microbial communities from these two locations were largely different and the species with different abundances,especially cyanobacteria,were found to be indicators that could reflect human activities and coral reefs' health *** dominant phyla (Proteobacteria,Firmicutes and Bacteroidetes) comprised 58.6% and 71.0% of the Pl and Gf communities,***,for Am,three dominant phyla(Proteobacteria,Firmicutes and Deinococcus-Thermus) comprised 97.3% of the communities and 2 predominant phylum(Proteobacteria and Cyanobacteria) comprised 80.95% of the A,B,C,D,E and H sites ***,the results of NMDS,PCoA and Clustering analysis showed that these samples could be divided into three groups:Am group,Moorea group and Pl+Gf group,and the results were consistent with the result of ***,based on Kruskal-Wallis test among classes and the pairwise Wilcoxon test amon

关键词： coral reefs microbial community symbiont cyanobacteria wellness indicators

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Effective Production of Dehydrogenation-Streptopyrrole from Streptomyces armeniacus CGMCC 7129 by Combination of Response Surface Methodology and Purification with High Performance Countercurrent Chromatography(HPCCC)

Effective Production of Dehydrogenation-Streptopyrrole from ...

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中国生物工程学会第六次全国会员代表大会暨第九届学术年会

作者：袁博 Chun-xia Yan Ye-Yan Qiu Sheng Qin Qiuyue Zhang Yun-xin Hao Xiu-Yun Ju Ji-Hong.Jiang 江苏师范大学江苏省药用植物生物技术重点实验室 State Key Laboratory of Plant Physiology and Biochemistry College of Biological SciencesChina Agricultural University Department of Biology and microbiology South Dakota State University The Key Laboratory of Biotechnology for Medicinal Plant of Jiangsu Provice Jiangsu Normal University School of Biology Science Jiangsu Normal University

Dehydro-streptopyrrole（DS） is a new class chemical of dehydrobenzoxazin that was first found in the endophytic streptomyces Streptomyces armeniacusi CGMCC *** has a more widely antimicrobial spectrum than ***,we found that metal ions(Pb,Mn,Cu and Cd) could improve the output of *** order to meet its sample supply for further in vivo pharmacological investigation,an efficient method was developed for the preparation of DS by a combination of response surface methodology（RSM） and preparative high performance liquid chromatography（PHPLC） from strain CGMCC *** heavy metal ion content in the culture was optimized by the RSM to enhance DS *** results revealed a model（P<0.0001） that was highly significant and this could consequently be utilized for optimizing *** maximum concentration of DS was measured at 800.00,382.31,599.97 and 600.00 μM levels of Pb,Mn,Cu and Cd,*** the optimized fermentation conditions,DS production was 75.36 μg/mL compared to 17.44 μg/mL under the original standard fermentation ***,a rapid and efficient method for the separation and purification of DS from crude culture extract was developed using high performance countercurrent chromatography（HPCCC）.The liquid phase solvent system employed for HPCCC separation was composed of nhexane-ethyl acete-methanol-water（5:7:6:5,v/v）.The isolation was accomplished within 140 min and the significantly purity of DS was over 97%.Collectively,we have therefore established the new technique of production and fast-separation of DS.

关键词： Streptomyces armeniacus Dehydrogenation-streptopyrrole(DS) Response surface methodology High performance countercurrent chromatography

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Gene cloning,expression and characterization of an exo-chitinase with high β-glucanase activity from Aeromonas veronii B565

Gene cloning,expression and characterization of an exo-chiti...

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中国生物工程学会第六次全国会员代表大会暨第九届学术年会

作者：霍凤敏冉超 Yalin Yang Jun Hu Zhigang Zhou 首都医科大学附属北京胸科医院中国农业科学院饲料研究所 Key Laboratory for Feed Biotechnology of the Ministry of Agriculture Feed Research InstituteChinese Academy of Agricultural Sciences

Chitinases（EC 3.2.1.14） are the primary enzymes that catalyze the hydrolysis of recalcitrant chitin into valuable soluble *** have a very broad range of applications in ***,the use of chitinase in aquafeed has rarely been *** veronii strain B565 is a gram-negative bacterium isolated from aquaculture pond sediment in China,and was found to have the ability to produce chitinase to control fungal or Myxozoa-related *** B565 encodes five chitinases,namely Chi92（AEB48885.1）,chitinase A（AEB48887.1）,ChiB565（AEB48892.1）,and two other chitinases（AEB48889.1,AEB50059.1）.In this study,the chitinase gene chi92 was cloned from *** strain B565 and expressed in Pichia pastoris *** recombinant chitinase（Chi92） was purified and the biochemical properties were ***92 was supplemented in diets containing *** powder and fed to zebrafish for 14 *** comparing with the control group,effect of Chi92 supplementation on growth,feed utilization,microvilli morphology,and disease resistance was *** complete gene sequence encoded a polypeptide with 864 amino *** sequence alignment indicated that Chi92 contained the consensus substrate binding sequence SXGG（Ser270-Val271-Gly272-Gly273） and catalytic sequence DXXDXDXE（Asp307-Gly308-Val309-Asp310-Ile311-Asp312-Trp313-Glu314） of family 18 ***92 was successfully expressed in *** with high chitinase activity（69.4 U/mL）.Chi92 exhibited optimal activity at pH 6.0 and 40 °***92 was resistant to protease trypsin,or pepsin and sensitive to chymotrypsin or tilapia intestinal *** Chi92 activity was increased by Mn2+（5 mmol/L） and mercaptoethanol（1 mmol/L）,and strongly inhibited by Cr,Cu,and Fe（1 mmol/L or 5 mmol/L）,and EDTA（5 mmol/L）.Thin-layer chromatography and electrospray ionization–coupled mass spectrometry revealed that N-diacetylglucosamine was the dominant product of Chi92 when colloidal chitin was

关键词： Chitinase Chi92 Aeromonas veronii B565 Zebrafish

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