拟南芥AtMYB44与RCAR1相互作用的研究

以拟南芥脱落酸(ABA)受体RCAR1和以文为诱饵蛋白质,利用酵母双杂交(Y2H)技术筛选出转录因子AtMYB44,本文进行了pull down及重转酵母验证,确定它们之间的关系,实验结果显示RCAR1与AtMYB44之间确实存在相互作用.此外,为了确定RCAR1在AtMYB44上的作用区域,构建了不同长度的AtMYB44cDNA序列的PGADT7载体,利用酵母双杂交实验分析,表明只有AtMYB44的N端54~105氨基酸序列与RCAR1存在相互作用.     

VDAC3与ABA受体蛋白相互作用的验证以及初步研究

酵母双杂交技术在拟南芥cDNA文库中筛选得到了与ABA受体蛋白RCAR1,RCAR3均有相互作用的蛋白质VDAC3,并已获得VDAC3基因的全长.VDAC3蛋白包含有3个结构域,分别为Porin3结构域,DUF3442结构域和SEG片段,现根据结构域分布,将VDAC3截短为包含DUF3442的VDAC3N以及包含SEG的VDAC3C两段.将VDAC3全长以及分别截短的169个氨基酸(VDAC3N)及105个氨基酸(VDAC3C)的片段进行酵母转化实验,与RCAR1或RCAR3共同转化后,VDAC3,VDAC3N的共转酵母能够在缺陷型培养基上生长,并且使X-Gal显色为蓝色,而VDAC3C则不能.这验证了全长的VDAC3与RCAR1,RCAR3的存在相互作用,并发现决定VDAC3蛋白与RCAR1,RCAR3是相互作用的结构域位于N端的DUF3442结构域.     

AtMYB44与ABI1竞争性结合ABA受体RCAR1的研究

为了进一步确定AtMYB44、ABI1、RCAR1三者之间的关系,本文进行了竞争性pull down和ABI1磷酸酶活性的测定实验,结果表明AtMYB44与ABI1能够竞争性结合RCAR1,并且这种竞争性结合依赖于ABA,AtMYB44能够降低RCAR1对ABI1活性的抑制,这为深入研究ABA信号转导途径奠定了基础.     

拟南芥CARK4与RCAR12相互作用的研究

本研究采用酵母双杂交(Y2H)、GST-融合蛋白沉降技术(GST-pull-down)及双分子荧光互补实验(BiFC)实验方法研究蛋白质激酶CARK4与脱落酸受体RCAR12之间的相互作用.酵母双杂交(Y2H)试验显示,缺失氮端的CARK4(CARK4ΔN)与RCAR12具有明显的相互作用.体外GST-融合蛋白沉降技术试验进一步得出CARK4能拉下RCAR12,而不能结合负对照GST,这证明CARK4可以直接与RCAR12相互作用.为了确定CARK4与RCAR12在植物体内是否也存在相互作用,利用农杆菌介导的烟草叶片瞬时表达来进行BiFC实验,结果表明在烟草中共注射CARK4与RCAR12能发激发荧光,而负对照却没有,这说明CARK4与RCAR12在植物体内存在相互作用.研究结果表明,CARK4与RCAR12在植物体内、体外均具有相互作用.     

ABA受体RCAR1/PYL9相互作用蛋白质的筛选及初步研究

使用拟南芥作为模型,采用酵母双杂交系统筛选出和ABA受体RCAR1/PYL9有相互作用的蛋白质,以更深入的研究RCAR1/PYL9的功能.首先利用反转录PCR方法获得拟南芥的cDNA,然后通过酵母双杂交系统,用拟南芥RCAR1/PYL9作为诱饵蛋白对拟南芥cDNA文库进行筛选,并获得多个阳性克隆.对该菌落进行进一步的鉴定得到与RCAR1/PYL9具有相互作用的蛋白质,这对进一步研究RCAR1/PYL9及其相互作用蛋白质在ABA信号转导途径中的作用及地位奠定了基础.     

拟南芥蛋白质激酶CARK7与ABA受体RCAR12相互作用的研究

脱落酸(ABA)受体RCARs在ABA信号传导过程中起关键作用,研究ABA受体的相互作用蛋白质对进一步了解植物抗逆的分子机制有重要的意义.本研究通过酵母双杂交发现CARK7与ABA受体RCAR12共转化酵母能在营养缺陷型平板SD/-Trp-Leu-Ade上生长.将CARK7与RCAR12分别克隆到原核表达载体pET-28a(+)和pGEX-6p-1中,构建融合蛋白表达载体,并进行原核表达和亲和纯化.纯化所得蛋白进行GST-pull down实验,结果表明RCAR12能够把CARK7蛋白拉下来.上述结果说明CARK7与RCAR12在体外存在相互作用.进一步通过双分子荧光互补实验发现CARK7与RCAR12在烟草叶片表皮细胞中能产生荧光信号,说明两者在体内同样具有明显的相互作用.     

拟南芥AtDWD与RCAR1相互作用的初步研究

采用pull down和酵母共转化的方法研究了AtDWD与RCAR1之间的相互作用.体外pull down试验得出AtDWD与RCAR1之间存在明显的相互作用;酵母共转化试验显示AtDWD的截断区域中只有WD45和WD345转化子可以在SD四缺培养基平板上长出正常的酵母菌落.实验结果表明,AtDWD C端的145~350位氨基酸序列中含有与RCAR1相互作用的功能域.     

酵母双杂交筛选与脱落酸受体相互作用的蛋白质及其重转酵母的验证

脱落酸（abscisic acid,ABA）是一种重要的植物激素,它能使植物适应逆境胁迫,从而调节植物的生长发育状况.现今ABA受体已被公认为RCARs/PYR/PYLs家族蛋白.本实验通过拟南芥为模式植物,利用酵母双杂交系统共转化方法从拟南芥cDNA文库中筛选与RCAR3相互作用的蛋白质,获得多个阳性克隆,从中选取一个与生长素调节相关的基因NIT1的全长cDNA重新转化酵母验证,发现与RCAR3仍然有相互作用,排除了假阳性的可能.从而为下一步研究该基因与RCAR3的相互关系做好了准备.     

拟南芥CARK3与RCAR12相互作用的研究

植物激素ABA是植物应对非生物胁迫的响应因子,广泛参与了植物生长发育的调控.ABA结合其受体后抑制PP2Cs的磷酸酶活性,释放SnRK2s,从而启动ABA信号转导途径.本文采用酵母双杂交系统,GST-pull down技术研究蛋白质激酶CARK3与ABA受体RCAR12在体外的相互作用,结果显示CARK3和RCAR12共转入酵母后,在三缺平板上能够正常生长;经His-Tag抗体检测,CARK3与RCAR12出现明显且单一的条带.同时,采用双分子荧光互补实验研究CARK3与RCAR12在体内的相互作用,结果显示CARK3与RCAR12共转化烟草后能观察到较强的荧光.体内外实验表明,CARK3与RCAR12存在明显的相互作用,CARK3可能通过直接磷酸化ABA受体RCAR12来调控ABA信号途径的生理应答.     

拟南芥E3泛素连接酶AtARRE与ABI5相互作用分析

为了阐述AtARRE与ABA信号通路关键转录因子ABI5的作用,本研究采用原生质体瞬间表达系统探究了AtARRE蛋白的亚细胞定位,证明AtARRE蛋白定位于细胞核.随后,采用酵母双杂交和GST-Pull down技术分析了AtARRE与ABI5在体外的相互作用,证明AtARRE与ABI5在体外存在相互作用.最后,本研究采用双分子荧光互补实验进一步分析AtARRE与ABI5在体内的相互作用,结果表明共表达AtARRE与ABI5在植物体内存在相互作用.这些结果共同表明AtARRE可能参与了ABI5介导的植物对逆境的响应.     

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.     

Design and analysis of generic LBS application developing platform

Location based services is promising due to its novel working style and contents.A software platform is proposed to provide application programs of typical location based services and support new applications developing efficiently. The analysis shows that this scheme is easy implemented, low cost and adapt to all kinds of mobile nework system.     

橡胶颗粒沥青混合料低温抗裂性试验研究

以AC-13级配为基础,将橡胶颗粒代替部分集料掺入混合料中,以低温弯曲试验为评价方法对不同橡胶颗粒掺量下沥青混合料的低温抗裂性进行研究,并引入应变能密度值对混合料的低温抗裂性进行综合评价.试验结果表明:橡胶颗粒沥青混合料试件的破坏微应变均超过2 300,满足冬寒区的技术指标;无论是否掺加橡胶颗粒,随着温度的下降,沥青混合料破坏时的最大弯拉强度增大,弯拉应变降低,劲度模量增大;弯曲应变能密度在胶粒掺量为1%左右时具有较大的弯曲应变能密度值,此时橡胶颗粒沥青混合料具有较好的低温抗裂性.     

Jane Eyre——A Feminist In The 19th Century

In the 19th century the society was controlled by men, and women were just appendants of them, they had not any rights and freedom. But Jane was an exception, she showed some characteristics of early feminist. Jane showed her characteristics of feminism in three aspects: rebellion, equality, and independence. These characteristics were helpful to her success, and feminism is the only way out for women of that time.     

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-