人角质细胞生长因子基因转化水稻的初探

角质细胞生长因子(Keratinocyte growth factor, KGF)在组织的损伤修复中起着重要的作用.在植物里面表达高活性的KGF具有成本低、易开发的优点.本实验将人角质细胞生长因子基因经农杆菌介导转入水稻中,获得110株再生植株.对再生植株进行GUS检测发现有44.7%植株呈阳性,PCR检测的结果也显示有52.6%的植株为阳性植株.     

人角质细胞生长因子-1在水稻中的表达

人角质细胞生长因子(Keratinocyte growth factor,KGF)在组织的损伤修复中起着重要的作用.植物细胞生物反应器是一种成本低、安全性好的蛋白生产系统.本研究将人KGF-1经农杆菌介导法转入水稻中,共获得38株抗性再生植株.Southern杂交和RT-PCR结果表明KGF-1基因整合入水稻基因组中且部分转化株中KGF-1基因得到转录.Western blotting检测结果显示有两株转化水稻中能够表达KGF-1.     

重组人角质细胞生长因子(KGF-2)在毕赤酵母中的克隆与表达

利用RT－PCR技术，从体外培养的人胚胎成纤维细胞中扩增出人角质细胞生长因子（KGF－2）的cDNA及基因序列，并将其基因序列克隆入质粒pGEM-T Easy中，经测序与文献报道序列一致，将KGF－2基因切出后定向插入酵母分泌性表达质粒pPICZαA中，转化毕赤酵母菌株GS115，获得的重组体经甲醇诱导可表达出人角质细胞生长因子KGF－2，表达的KGF－2经Ni^2 柱亲和层析纯化后，有很好的生物学活性。     

人角质细胞生长因子-1转化番茄的研究

人角质细胞生长因子在组织的损伤修复中起着重要的作用.以番茄子叶为外植体,通过根癌农杆菌介导法,将人KGF -1导入番茄中,共获得12株独立的抗性转化植株,PCR和DNA斑点杂交结果表明:KGF-1基因整合入番茄基因组中,为获得植物源表达的KGF -1蛋白打下了基础.     

人角质细胞生长因子的植物表达载体构建

人角质细胞生长因子(KGF)有3个外显子,2个内含子.以人DNA为模板,PCR扩增获得角质细胞生长因子外显子2和外显子3;合成单链的寡核苷酸,经链延伸和PCR扩增从而获得外显子1.应用延伸PCR将3个外显子连接得KGF编码序列,并连接CaMV35S启动子和NOS终止子后克隆至植物表达质粒pBI 1301.     

高密度培养大肠杆菌表达人角质细胞生长因子

角质细胞生长因子(KGF)是成纤维生长因子家族(FGF)成员,与上皮细胞增殖、组织胚胎发育、创伤修复等过程密切相关.在大肠杆菌BL21(DE3)中克隆KGF基因,并进行诱导表达条件的优化.使用合成培养基,应用底物反馈流加策略将糖浓度控制在较低水平,在发酵罐中高密度培养,细胞干质量浓度可以达到46 g/L.发酵罐培养的大肠杆菌在对数生长期经过异丙基-β-D-硫代半乳糖苷(IPTG)诱导,在可溶性和不溶性产物中都得到了具有免疫学活性的GST-KGF融合蛋白,经过ELISA检测,发酵液中可溶性表达的KGF产率达到108μg/L.该研究为利用原核表达系统生产活性KGF建立了优化工艺,有利于将大规模生产用于创伤修复的KGF.     

红隼肺的组织结构及相关活性物质表达

红隼(Falco tinnunculus)属国家级保护动物,研究其器官组织及相关活性物质的表达,对于动物学和保护生物学研究都有重要意义.为了搞清红隼肺的组织结构特征,观察角质细胞生长因子(KGF)、c-Fos蛋白和Bax蛋白在肺中的表达情况,应用生物显微技术和免疫组织化学方法观察了2只红隼肺的组织结构,检测了KGF、c-Fos和Bax在肺中的表达.结果表明,红隼肺的表面覆盖有浆膜,肺实质由各级支气管及其周围的呼吸毛细管组成,肺房呈辐射状开口于三级支气管周围,呼吸毛细管相互吻合成网状,周围有丰富的毛细血管网.KGF、c-Fos和Bax在三级支气管的平滑肌细胞、肺房和呼吸毛细管的上皮细胞中呈免疫反应阳性.KGF、c-Fos和Bax可能发挥着不同的功能,它们的协同表达可能调控着鸟肺细胞的生长、发育及成熟.     

重组人角质细胞生长因子2在伤口愈合及毛发再生方面的影响

探寻重组人角质细胞生长因子2(rh-KGF2)和易孚(EGF为主要的药品)对SD大鼠烫伤创面愈合方面效果的差异,同时探寻重组人角质细胞生长因子2(rhKGF2)和米诺地尔在毛发再生方面效果的差异.通过建立大鼠的自身对照模型,左侧形成深Ⅱ°烫伤创面,右侧涂抹脱毛膏脱毛.通过连续4~6周,每日1次对烫伤创面和脱毛区域采用涂抹的方式对伤口给药.在此过程中对大鼠的一般状态、体重、烫伤愈合效果以及新生毛长及毛重进行观察.通过分析得出的结论为:rh-KGF2在伤口愈合方面和毛发再生方面有良好的效果.     

农杆菌介导IPT和etr1-1双基因转化番茄

建立了番茄子叶高频再生体系，优化了农杆菌介导的外源基因转化番茄的条件，将IPt和etrl-1双基因导入番茄中，以除草剂PPT作为筛选标记，得到3株转化再生植株．通过PCR，RT-PCR检测证明etrl-1和IPT基因已整合到转基因番茄植株的基因组中，并在转录水平有一定表达．     

BYDV CP基因转化小麦原生质体及转基因植株再生

用原生质体融合技术与ＰＥＧ介导的直接转基因方法相结合，使高频率分裂的小麦济南１７７悬浮细胞系原生质体与具有分化能力的小麦济南１７７胚性意伤组织原生质体融合，形成具有分裂和分化能力的融合细胞；同时，利用ＰＥＧ对原生质体的直接转基因作用，将带有抗病毒基因（ＢＹＤＶＣＹｇｅｎｅ）的质粒Ｐｐｐ１５与带有抗性选择标记基因的质粒严ＰＢｌＡｃｔＳＮ导入融合细胞中．经抗性筛选获得抗潮霉素（Ｈｍ）的阳性克隆并再生植株．对再生植株作ＰＣＲ检测表明，ＣＰ基因已整合到转化植株的基因组中并稳定表达．     

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     

On the Thermodynamic Limit for Responses to Static Electromagnetic Fields

For molecular systems,the quantum-mechanical treatment of their responses to static electromagnetic fields usually employs a scalar-potential treatment of the electric field and a vector-potential treatment of the magnetic field.Although the potential for each field separately is associated with the choice of an(unphysical)origin,the precise choice of the origin for the electrostatic field has little consequences for the results.This is different for the