农杆菌介导的DREB1A基因转化多花黑麦草及其转化体系的优化

采用携带卡那霉素抗性基因nptII和GUS基因的ubiquitin启动子驱动的表达载体pBI121／DREB1A的根癌农杆菌AGL1, 对多花黑麦草幼胚来源的胚性愈伤组织进行了遗传转化,并优化了各种影响因素。胚性愈伤组织经根癌农杆菌感染和共培养后,用50mg／L巴龙霉素筛选抗性愈伤组织,待抗性愈伤组织在IB分化培养基上分化成苗后用25mg／L卡那霉素进一步筛选再生植株, 获得了部分抗性植株。抗性植株的总DNA用DREB1A基因的特异引物进行PCR检测,转化频率为2.14％,PCR-Southern blot进一步验证了转化植株基因组中含有该外源基因。各种影响转化效率因素的优化实验表明,当转化时菌液浓度的OD600为2.0、侵染时间为1h、共培养时间为2d、共培养温度为21℃及在共培养期间使用乙酰丁香酮等,均可明显提高转化频率。     

根癌农杆菌介导的骏枣愈伤组织遗传转化体系的研究

采用农杆菌介导法,对骏枣愈伤组织遗传转化体系进行研究.结果表明：进行遗传转化的合适条件为：愈伤诱导培养基MS＋6-BA 0.4 mg/L＋2,4-D 1.2 mg/L;浸染之前对愈伤组织进行6d的避光培养可提高愈伤组织在共培养过程中的成活率;愈伤组织在OD600=0.4～0.6的农杆菌菌液中浸染10 min,农杆菌LBA4404的生长情况最好;在28℃黑暗条件下共培养放置16 d对转化最适宜;头孢霉素的抑菌浓度为250 mg/L;卡那霉素的筛选浓度为125 mg/L.经PCR检测,初步证明外源目的基因已整合到骏枣基因组中,初步实现了农杆菌介导的骏枣愈伤组织遗传转化,与以骏枣茎叶外植体建立的遗传转化体系相比,以骏枣愈伤组织为外植体共培养时间长,有利于转化.     

水仙胚性愈伤的获得及农杆菌介导GUS基因的遗传转化

用正交实验设计对前期影响水仙胚性愈伤诱导和分化关联性大的因素进行综合分析,结果显示,培养基类型、激素及处理方式、不同添加物等对水仙外植体胚性愈伤诱导及分化的影响是基于培养基的多因素协同效应。水仙适宜胚性愈伤诱导培养基是:改良MB+NAA(0.2 mg/L)+6BA(2 mg/L)+ABA (2 mg/L)+AgNO3(5 mg/L);适宜的分化培养基是：改良N6+2,4D(0.5 mg/L)+6BA(2.5 mg/L)+NAA(0.2 mg/L)+ABA(2 mg/L)。实验中观察到水仙体细胞胚的发生,在胚性愈伤诱导和分化体系优化基础上实现农杆菌介导GUS基因转化,稳定表达抗性愈伤GUS染色显示最佳胚性愈伤诱导培养基能促进水仙外植体感受态的形成,有利于外源基因的接受,PCR检测表明实验中获得3株转GUS基因阳性植株。     

口蹄疫抗原决定簇融合基因转化胡萝卜的研究

利用转基因植物生产疫苗是目前植物基因工程的一个研究热点,具有生产简单、成本较低、使用安全方便、易贮存等优点.本实验通过农杆菌介导的遗传转化,以O型和A型口蹄疫抗原决定簇基因O21-O14-A21转化胡萝卜.通过筛选,获得潮霉素抗性愈伤组织,经胚状体再生得到156棵抗性苗.通过GUS染色检测,GUS报告基因在部分转化的愈伤组织中瞬时表达,并在部分抗性苗中稳定表达.对部分抗性植株进行PCR和PCR-Southern杂交检测,证实目的基因已经成功整合到胡萝卜基因组中.     

胡萝卜下胚轴遗传转化体系的优化

【研究目的】优化胡萝卜下胚轴转化体系,为胡萝卜基因工程研究提供一个简便而快捷的方法;【方法】以胡萝卜下胚轴为外植体,对农杆菌介导的的遗传转化体系进行优化;【结果】实验结果表明:1)将MS培养基的有机成分进行调整配制成基本培养基MST,有利于愈伤组织的诱导,但不利于苗的分化与再生培养;2)农杆菌侵染外植体的时间为15 min,共培养时间为50 h,获得抗性愈伤组织诱导率达78%;3)羧苄青霉素的浓度从愈伤组织诱导阶段的500 mg/L下降到苗再生阶段的100 mg/L时,提高抗性苗再生率到88%;【结论】通过体系的优化,极大的提高了胡萝卜下胚轴的转化效率,为胡萝卜基因工程的研究提供有效的技术平台。     

口蹄疫抗原决定簇融合基因转化胡萝卜的研究

利用转基因植物生产疫苗是目前植物基因工程的一个研究热点，具有生产简单、成本较低、使用安全方便、易贮存等优点．本实验通过农杆菌介导的遗传转化．以O型和A型口蹄疫抗原决定簇基因O21-O14-A21转化胡萝卜．通过筛选．获得潮霉素抗性愈伤组织。经胚状体再生得到156棵抗性苗．通过GUS染色检测．GUS报告基因在部分转化的愈伤组织中瞬时表达．并在部分抗性苗中稳定表达．对部分抗性植株进行PCR和PCR—Southern杂交检测．证实目的基因已经成功整合到胡萝卜基因组中．     

农杆菌介导的花生遗传转化体系的初步研究

应用花生胚小叶再生体系,以农杆菌介导法对质粒上的基因转化进行初步探讨.通过对影响基因转化的各因素,如抗生素的筛选压、共培养时间、AS的影响、及外植体取材方式等进行研究,结果发现:外植体与农杆菌在加有AS(100μmol/L)的改良MS液体培养基中共培养4 d后,先进行10 d的恢复培养,继而转入加有Hy(25 mg/L)和Cef(200 mg/L)的抗性筛选培养基中进行筛选培养40～50 d,后转入含Hy(20 mg/L)的分化培养基中分化,3个月后在首批材料中得到6个再生植株,经PCR检测,有1株显示了800 bp的GUS基因核酸片段,初步证实了此转化体系合理有效.     

刺五加愈伤组织的遗传转化

刺五加胚性愈伤组织经根癌农杆菌Ｃ５８Ｃ１感染后，共培养４７－７２ｈ，再转移到含氯霉素５０ｍｇ／Ｌ的ＭＳ选择培养基上继代培养５０－６０ｄ，筛选得到氯霉素抗性愈伤组织。该愈伤组织能在无激素的ＭＳ培养基上增殖，并检测到胭脂碱合成酶活性，表明是转化愈伤组织。     

银合欢壳多糖酶在转基因Khao Da wk Mali 105籼稻中的表达(英文)

通过含有pCAMBIA1300载体(抗潮霉素基因(hpt)作为选择标记基因)的根癌农杆菌菌株EHA105的介导,将326个银合欢I型碱性壳多糖酶基因(KB3,GenBank登录号为:AAM49597)的氨基酸导入到籼稻Khao Dawk Mali 105(KDML 105)植株中,获得了KDML 105籼稻的愈伤组织。pCAMBIA1300载体包含有CaMV35S启动子、银合欢壳多糖酶基因和NOS终止子。在NB培养基的水稻秧苗中愈伤组织的转化率为94%.在培养温度为28℃,以含有4 mg/l BAP和7 g/l植物凝胶再生培养基的条件下,愈伤组织的再生频率高达80%.为了建立一个KDML 105农杆菌转化方法,考察了各方面的因素。共培养期间,在培养基中添加50μM的乙酰丁香酮对提高瞬时转化率非常重要。4周龄的盾片衍生的愈伤组织是最佳的起始材料。添加40 mg/l潮霉素和400 mg/l头孢霉素的NB培养基是水稻愈伤组织选择性转化的最佳培养基,其转化率达到78%.采用PCR、Northern Blot杂交凝胶、Southern Blot杂交凝胶和GUS分析技术研究了愈伤组织和转基因水稻中的壳多糖酶和gus基因的表达。壳多糖酶和gus基因在水稻的多个部位被检测到,包括愈伤组织、叶、根、茎和秧苗,但是它们在水稻根部的含量较少。抗真菌的水稻壳多糖酶表现出抗串珠镰刀菌的性质。     

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%.     

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.     

Characterization of Ordovician carbonate reservoirs, southeastern Saskatchewan, Canada

The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3～4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰～ -7.8‰, PDB) and matrix dolomites (-6.6‰～ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084～0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078～0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.     

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.     

A computer generator for randomly layered structures

AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…     

An Index mechanism for multi-scale view in spatial databases

There are numerous geometric objects stored in the spatial databases. An importance function in a spatial database is that users can browse the geometric objects as a map efficiently. Thus the spatial database should display the geometric objects users concern about swiftly onto the display window. This process includes two operations:retrieve data from database and then draw them onto screen. Accordingly, to improve the efficiency, we should try to reduce time of both retrieving object and displaying them. The former can be achieved with the aid of spatial index such as R-tree, the latter require to simplify the objects. Simplification means that objects are shown with sufficient but not with unnecessary detail which depend on the scale of browse. So the major problem is how to retrieve data at different detail level efficiently. This paper introduces the implementation of a multi-scale index in the spatial database SISP (Spatial Information Shared Platform) which is generalized from R-tree. The difference between the generalization and the R-tree lies on two facets: One is that every node and geometric object in the generalization is assigned with a importance value which denote the importance of them, and every vertex in the objects are assigned with a importance value,too. The importance value can be use to decide which data should be retrieve from disk in a query. The other difference is that geometric objects in the generalization are divided into one or more sub-blocks, and vertexes are total ordered by their importance value. With the help of the generalized R-tree, one can easily retrieve data at different detail levels.Some experiments are performed on real-life data to evaluate the performance of solutions that separately use normal spatial index and multi-scale spatial index. The results show that the solution using multi-scale index in SISP is satisfying.     

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-     

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-