短小芽孢杆菌基因无痕修饰系统的建立及应用

为了实现对短小芽孢杆菌(Bacillus pumilus)SCU11基因组连续地无痕改造,提高碱性蛋白酶产量,本文建立了基于Upp基因反向筛选的短小芽孢杆菌基因无痕修饰系统,将碱性蛋白酶基因AprE的1份拷贝无痕插入至短小芽孢杆菌染色体16S rDNA区.摇瓶发酵实验显示,突变菌株碱性蛋白酶活最高达到7125 U/mL,与出发菌株相比提高了33.1%.结果表明利用该系统可实现对短小芽孢杆菌基因组的无痕修饰,对AprE插入突变菌株进行双交换筛选的最终效率约为23.07%.     

短小芽孢杆菌sRNA Bpsr112的鉴定及功能研究

为了给短小芽孢杆菌的改造提供新思路,提高碱性蛋白酶产量,实验室前期对短小芽孢杆菌进行了转录组测序,预测了短小芽孢杆菌的sRNA.本研究通过生物信息学方法和Northern杂交鉴定了一个新的sRNA Bpsr112.然后构建了Bpsr112的敲除型菌株和过表达菌株,利用相关菌株进行了生长曲线、盐胁迫和蛋白酶活等实验,再利用SDS-PAGE检测胞外蛋白酶AprE的表达水平.结果表明,在发酵至60 h和72 h时,与对照菌株相比,敲除型菌株酶活显著降低（P<0.01）,过表达菌株酶活显著提高（P<0.01）;同时SDS-PAGE结果表明,AprE蛋白含量在敲除菌株中降低,在过表达菌株中提高,说明sRNA Bpsr112对短小芽孢杆菌蛋白酶活具有正调控.本研究鉴定了短小芽孢杆菌中的sRNA,并发现Bpsr112对蛋白酶活具有促进作用.     

绿色荧光蛋白(GFP)基因在短小芽孢杆菌中的组成型表达

采用PCR技术,亚克隆来自短小芽孢杆菌总基因组中的启动子活性片段F1,构建了一个大肠杆菌-短小芽孢杆菌穿梭表达载体pHY300-F1gfp,以缺失启动子的绿色荧光蛋白(GFP)基因为报告基因,检测了该片段在短小芽孢杆菌DX01菌株中的启动活性,在荧光显微镜下,观察到了明亮的绿色荧光,证实活性片段F1具有组成型启动子的功能,GFP基因在短小芽孢杆菌中实现了组成型表达．     

含Cry1Ab/Ac基因的白僵菌工程菌的构建

以来源于苏云金芽孢杆菌的Bt-Cry1Ab/Ac基因为目的基因,以潮霉素Hyg为抗性标记基因,以绿色荧光蛋白基因GFP为报告基因的真菌表达载体,采用农杆菌介导真菌遗传转化(ATMT)法,将Cry1Ab/Ac基因插入白僵菌基因组中,先后用潮霉素B抗性筛选、荧光检测、真菌基因组PCR分子检测法、Bt-Cry1Ab/Ac基因检测试纸条多种方法进行转基因工程菌株鉴定,结果证实了目的基因成功插入白僵菌基因组,为白僵菌杀虫活性的深入研究奠定了基础.     

植物乳杆菌CS3全基因组分析及对己醛的转化

大豆是传统酱油酿造的主要原料,在酿造过程中容易发生氧化变质和酸败等现象,从而产生不良风味和有害物质,其原因是发酵过程中产生了不饱和脂肪醛类物质。本研究以植物乳杆菌(Lactiplantibacillusplantarum)CS3为研究对象,在对该菌株进行全基因组测序和己醛胁迫下的转录组分析的基础上,筛选出转化己醛的关键基因AdhE(双功能乙醛CoA/酒精脱氢酶)和己醛胁迫下的高表达转录因子Fru R。利用CRISPR-Cas9基因编辑技术,将基因AdhE和FruR进行基因敲除验证,敲除后的菌株在MRS培养基中代谢己醛的能力分别下降了17.72%和56.72%。这表明两个基因在己醛转化过程中都发挥重要作用。这一结果为植物乳杆菌在酱油生产中降低醛类等有害物质的应用提供了理论支持。     

核盘菌交配型基因mat1-1敲除载体的构建及转化

利用根癌农杆菌介导真菌遗传转化方法对核盘菌的交配型基因mat1-1进行基因同源重组的敲除对比实验,获得了缺失mat1-1基因的转化菌株.先利用PCR方法获得mat1-1基因的左右两侧片段,将测序正确的两个侧翼片段分别重组到农杆菌转化载体PBI-G3C中,并将新霉素抗性标记引入农杆菌转化载体PBI-G3C中,构建成农杆菌介导转化核盘菌的打靶载体ΔPBI-G3CN-mat1-1,再将ΔPBI-G3CN-mat1-1质粒转化至根癌农杆菌EHA105中.利用核盘菌菌丝进行转化,将得到的转化菌株,利用PCR方法进行验证,证实有敲除菌株存在.通过对敲除菌株进行生理表型的测定发现,缺失mat1-1基因的敲除菌株其生长速度与野生核盘菌菌株相比无明显差异,但敲除菌株不能产生菌核与子囊盘.     

一株短小芽孢杆菌的16S rDNA基因序列分析

从黄颡鱼中分离得到一株优势菌BP-1,经过形态观察、生理生化试验和16S rDNA基因序列分析,表明该菌为发酵型,能运动,革兰氏阳性杆菌.获得的16S rDNA基因序列长度为1478 bp,Gen Bank数据库中登录号为KP299290.该序列与GenBank数据库中多条短小芽孢杆菌16S rDNA基因序列相似性高达99%,系统进化树上与短小芽孢杆菌亲缘关系最近,从而判定菌株BP-1为短小芽孢杆菌.这为短小芽孢杆菌的鉴定和分类提供科学依据.     

根癌农杆菌介导的丝状真菌简青霉遗传转化的研究

利用根癌农杆菌LBA4404介导,建立了丝状真菌简青霉(Penicillium simpli-cissimum)H5的遗传转化系统.潮霉素抗性筛选、PCR和Southern blot分子鉴定等结果表明:筛选获得的转化子能够稳定遗传,外源的T-DNA以单拷贝随机整合到简青霉的基因组中.实验初步研究了农杆菌浓度、乙酰丁香酮浓度和共培养时间等因素对转化效率的影响,经过优化后转化体系的效率可达50个转化子/105个孢子.农杆菌介导的遗传转化方法在简青霉上的运用,将为研究该菌的基因工程改造提供强有力的工具.     

农杆菌介导东方百合“西伯利亚”遗传转化体系建立

以东方百合"西伯利亚"(Lilium seberia)为材料,建立了百合再生和农杆菌介导的遗传转化体系.采用LBA4404和GV3101菌株对鳞片来源的愈伤组织进行侵染,并以鳞片作为对比转化材料,两种菌株携带有相同的双元载体pCAMBIA1301.研究结果表明,农杆菌侵染愈伤组织外植体转化率较高;乙酰丁香酮(AS)的添加能大大提高根癌农杆菌转化百合的效率.另外,共培养时间、不同菌株均对转化效率有一定的影响.筛选后获得的抗性植株经PCR检测和GUS组织化学染色证实外源基因已经整合到基因组DNA中并获得表达,转化率为5%～8%.     

芽孢缺失对D-核糖产量的影响

利用原生质体紫外线诱变的方法处理转酮酶缺陷型短小芽孢杆菌,选育到一株丧失芽孢形成能力的转酮酶缺陷型短小芽孢杆菌,摇瓶培养D-核糖平均产量达到66g/L,较出发菌株提高38%.研究表明,芽孢生成缺陷有利于D-核糖的产生,筛选芽孢生成能力缺陷的菌株是选育D-核糖高产菌株的有效手段.     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

An Analysis of Catherine＇s Character——Wuthering Heights

Wuthering Heights, Emily Bronte＇s only novel, was published in December of 1847 under the pseudonym Ellis Bell. The book did not gain immediate success, but it is now thought one of the finest novels in the English language. Catherine is the key character of this masterpiece, because everybody and everything center on her though she had a short life. We can understand this masterpiece better if we know Catherine well.     

Lower Paleozoic oil relationships within Williston Basin, Canada

The Williston Basin is a significant petroleum province, containing oil production zones that include the Middle Cambrian to Lower Ordovician, Upper Ordovician, Middle Devonian, Upper Devonian and Mississippian and within the Jurassic and Cretaceous. The oils of the Williston Basin exhibit a wide range of geochemical characteristics defined as "oil families", although the geochemical signature of the Cambrian Deadwood Formation and Lower Ordovician Winnipeg reservoired oils does not match any "oil family". Despite their close stratigraphic proximity, it is evident that the oils of the Lower Palaeozoic within the Williston Basin are distinct. This suggests the presence of a new "oil family" within the Williston Basin. Diagnostic geochemical signatures occur in the gasoline range chromatograms, within saturate fraction gas chromatograms and biomarker fingerprints. However, some of the established criteria and cross-plots that are currently used to segregate oils into distinct genetic families within the basin do not always meet with success, particularly when applied to the Lower Palaeozoic oils of the Deadwood and Winnipeg Formation.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that     

Call for Papers

<正>"The Journal of Shanghai Normal University:Mathematics"is published by Shanghai Normal University as regular issues of The Journal of Shanghai Normal University each year from 2014 in English.The editors-in-chief of the issues are professors Yuhao Cong and Maoan Han.The Journal of Shanghai Normal University was started in 1958 with