Erwinia persicina is a bacterium that has been known to produce secondary metabolites,such as andrimid,pink pigment,and exopolysaccharides,and to infect more than twenty plant species.However,traditional gene manipula...Erwinia persicina is a bacterium that has been known to produce secondary metabolites,such as andrimid,pink pigment,and exopolysaccharides,and to infect more than twenty plant species.However,traditional gene manipulation methods have been hindered by the inefficient of suicide plasmid-mediated genome editing.In this study,we describe the successful application of the CRISPR-Cas9 system in E.persicina.Efficient genome editing was achieved by substituting the native gRNA promoter with J23119 in a single-plasmid system(pRed_Cas9_ΔpoxB)and optimizing the gRNA design.The use of double gRNAs led to the deletion of a 42 kb genomic fragment,and the incorporation of a sacB screening marker facilitated iterative knockouts.Additionally,a 22 kb plasmid containing a self-resistance gene was conjugally transferred into E.persicina,resulting in the insertion of a 6.4 kb fragment with 100%efficiency.Furthermore,we demonstrated the expression of shinorine,an anti-UV compound,within the E.persicina chassis.This study establishes E.persicina as a promising chassis for synthetic biology and provides a model for gene-editing systems in non-model microorganisms.展开更多
针对预热压力容器罐出料口堵塞、颗粒预热不足等性能缺陷,提出融合TRIZ(theory of inventive problem solving)创新理论与数值模拟的系统解决方案。通过构建技术系统功能模型、专创融合解题流程及性能提升框架,结合ARIZ算法生成多方案...针对预热压力容器罐出料口堵塞、颗粒预热不足等性能缺陷,提出融合TRIZ(theory of inventive problem solving)创新理论与数值模拟的系统解决方案。通过构建技术系统功能模型、专创融合解题流程及性能提升框架,结合ARIZ算法生成多方案并优选“设置分流锥”方案。基于颗粒预热数学模型与气固耦合模型,采用ANSYS CFX对60°、90°、120°顶角分流锥的流道工况进行数值模拟,对比分析流速、湍流涡耗散、湍流动能等流场特征,并通过Q-criterion准则解析旋涡结构演化规律。结果表明:上部锥形顶角60°分流锥方案最优,可显著提升流动与预热性能,降低功能退化量;有限单元法验证显示其结构可靠性满足设计要求。可见,提出的方案可谓探索预热压力罐优化设计提供新思路。展开更多
Ciliates are one of the oldest living eukaryotic unicellular organisms, widely distributed in the waters around the world. As a typical marine oligotrich ciliate, Strombidium sulcatum plays an important role in marine...Ciliates are one of the oldest living eukaryotic unicellular organisms, widely distributed in the waters around the world. As a typical marine oligotrich ciliate, Strombidium sulcatum plays an important role in marine food webs and energy flow. Here we report the first deep se- quencing and analyses of RNA-Seq data from Strombidium sulcatum. We generated 42,640 unigenes with an N50 of 1,451 bp after de novo assembly and removing rRNA, mitochondrial and bacteria contaminants. We employed SPOCS to detect orthologs from S. sulcatum and 17 other ciliates, and then carried out the phyloge- nomic reconstruction using 127 single copy orthologs. In phylogenomic analyses, concatenated trees have similar topological structures with concordance tree on the class level. Together with phylogenetic networks analysis, it aroused more doubts about the placement of Protocruzia, Mesodinium and Myrionecta. While epi- plasmic proteins are known to be related to morphological characteristics, we found the potential relationship between gene expression of epiplasmic proteins and morphological characteristics. This work supports the use of high throughput approaches for phylogenomic analysis as well as correlation analysis between expression level of target genes and morphological characteristics.展开更多
基金funding from the Hundred Talents Program of the Chinese Academy of Sciences to LZ(E3J56201).
文摘Erwinia persicina is a bacterium that has been known to produce secondary metabolites,such as andrimid,pink pigment,and exopolysaccharides,and to infect more than twenty plant species.However,traditional gene manipulation methods have been hindered by the inefficient of suicide plasmid-mediated genome editing.In this study,we describe the successful application of the CRISPR-Cas9 system in E.persicina.Efficient genome editing was achieved by substituting the native gRNA promoter with J23119 in a single-plasmid system(pRed_Cas9_ΔpoxB)and optimizing the gRNA design.The use of double gRNAs led to the deletion of a 42 kb genomic fragment,and the incorporation of a sacB screening marker facilitated iterative knockouts.Additionally,a 22 kb plasmid containing a self-resistance gene was conjugally transferred into E.persicina,resulting in the insertion of a 6.4 kb fragment with 100%efficiency.Furthermore,we demonstrated the expression of shinorine,an anti-UV compound,within the E.persicina chassis.This study establishes E.persicina as a promising chassis for synthetic biology and provides a model for gene-editing systems in non-model microorganisms.
文摘针对预热压力容器罐出料口堵塞、颗粒预热不足等性能缺陷,提出融合TRIZ(theory of inventive problem solving)创新理论与数值模拟的系统解决方案。通过构建技术系统功能模型、专创融合解题流程及性能提升框架,结合ARIZ算法生成多方案并优选“设置分流锥”方案。基于颗粒预热数学模型与气固耦合模型,采用ANSYS CFX对60°、90°、120°顶角分流锥的流道工况进行数值模拟,对比分析流速、湍流涡耗散、湍流动能等流场特征,并通过Q-criterion准则解析旋涡结构演化规律。结果表明:上部锥形顶角60°分流锥方案最优,可显著提升流动与预热性能,降低功能退化量;有限单元法验证显示其结构可靠性满足设计要求。可见,提出的方案可谓探索预热压力罐优化设计提供新思路。
基金This work was supported by the Natural Science Foundation of China （Grant Nos. 31430077, 91131013, and 31272285）. Many thanks are given to Prof. Weibo Song （Ocean University of China）, for his indispensable advice during the preparation of the manuscript.
文摘Ciliates are one of the oldest living eukaryotic unicellular organisms, widely distributed in the waters around the world. As a typical marine oligotrich ciliate, Strombidium sulcatum plays an important role in marine food webs and energy flow. Here we report the first deep se- quencing and analyses of RNA-Seq data from Strombidium sulcatum. We generated 42,640 unigenes with an N50 of 1,451 bp after de novo assembly and removing rRNA, mitochondrial and bacteria contaminants. We employed SPOCS to detect orthologs from S. sulcatum and 17 other ciliates, and then carried out the phyloge- nomic reconstruction using 127 single copy orthologs. In phylogenomic analyses, concatenated trees have similar topological structures with concordance tree on the class level. Together with phylogenetic networks analysis, it aroused more doubts about the placement of Protocruzia, Mesodinium and Myrionecta. While epi- plasmic proteins are known to be related to morphological characteristics, we found the potential relationship between gene expression of epiplasmic proteins and morphological characteristics. This work supports the use of high throughput approaches for phylogenomic analysis as well as correlation analysis between expression level of target genes and morphological characteristics.
