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1.
用设计合成的一对特异性引物,以马铃薯卷叶病毒中国株PLRV—Ch的RNA为模板,经反转录PCR扩增,将获得ORF2a的5′端0.6kb cDNA克隆于pUC19中,构建成重组质粒pLR8.PCR检测和酶切检测表明,获得了ORF2a的5′端0.6kb cDNA克隆,从该cDNA两端进行了两次序列分析,并将获得的核苷酸序列与国外报道的4个PLRV分离株的相应区域的同源性进行了比较,5个序列间具高度同源性。  相似文献   

2.
用设计合成的一对特异性引物,以马铃薯卷叶病毒中国株PLRV-Ch的RNA为模板,经反转录PCR扩增,将获得的ORF2a的3′端1kb cDNA克隆于pUCl9中,构建成重组质粒pLR7,经PCR检测、酶切检测,表明获得了ORF2a的3′端1kb cDNA克隆.从两端进行了两次序列分析,将获得的核苷酸序列与国外报道的四个PLRV分离株的相应区域的同源性进行了比较.结果表明它们具有高度同源性.文中对核苷酸中存在的可能移码序列及其下游的茎环结构(或假节结构),以及上游特征性三次重复序列进行了分析和讨论,发现上游特征性三次重复序列连续排列可形成几个连续的互补双链区和发夹结构,这一结构可能与移码有关.  相似文献   

3.
用聚合酶链式反应(PCR)技术,从新疆绵羊全血中扩增了β-乳球蛋白5′端调控区890bp的片段.通过T4 DNA连接酶将其连接于质粒载体pGEM-T上,转化至受体菌DH5α中.提取质粒经PCR扩增、酶切鉴定,证明重组质粒pT-BLG中含有β-乳球蛋白5′端调控区基因片段.经核苷酸序列测定,克隆的片段与发表的基因序列高度一致.  相似文献   

4.
酵母PHO81基因是调控阻遏酸性磷酸酯酶的的一种中介因子,其编码区由3531个核苷酸组成。由于片段较大,目前很难应用PCR技术一次予以扩增。本文用自行设计的4个引物以啤酒酵母总染色体DNA为模板借助PCR技术扩增PHO81基因的两个片段,其长度分别为1.3kb和2.2kb,通过限制性酶切将其装入载体质粒pUC18中。并初步进行了重组质粒限制性内切酶酶谱分析和Southern杂交试验。  相似文献   

5.
大肠杆菌F107菌毛A亚单位基因的克隆与鉴定   总被引:6,自引:0,他引:6  
用PCR技术,从水肿病大肠杆菌临床分离株中扩增出F107A亚单位基因(fedA)的510bp的全序列。将该PCR扩增产物在BamHⅠ和EcoR Ⅰ位点克隆进pUC18质粒载体,并转化大肠杆菌TG1,再根据限制性内切酶酶切分析筛选出含有fedA的重组质粒pf107G。将重组质粒进行序列测定,结果表明,此重组质粒中的插入序列与发表的fedA是一致的,证明该重组质粒就是含有fedA基因的重组质粒。  相似文献   

6.
将本室克隆的编码复制酶基因3′端约1/2序列及其3′端非编码区的苜蓿花叶病毒中国分离株(AIMV-Ch)RNA:3′端cDNA,重组到植物表达载体pROKⅡ中,通过致瘤农杆菌(Agrobacterium tumefaciens)介导,以叶圆片为转化材料,转化普通烟草,并获得了转基因植株.经卡那霉素抗性选择、PCR检测目的基因证明.AIMV RNA:3′端基因已整合到转基因烟草的基因组DNA中.转基因植物的攻毒试验表明转基因植株对苜蓿花叶病毒产生高水平的抗性.  相似文献   

7.
甜瓜多聚半乳糖醛酸酶基因cDNA的克隆和序列分析   总被引:3,自引:0,他引:3  
以甜瓜品种河套蜜瓜成熟果实mRNA为模板,经反转录合成和PCR扩增得到编码多聚半乳糖醛酸酶(Polygalacturonase,PG)基因全长cDNA,将其克隆于pUC19质粒中获得重组质粒pCMPG.此cDNA长1183bp,包括一个393个氨基酸残基组成的开放阅读框架,与已报道的甜瓜PG基因cDNA核苷酸序列比较同源性为99.3%,相应的氨基酸的同源性为98.5%.  相似文献   

8.
根据国外报道的 JEV( Japanese encephalitis virus)基因组全序列 ,针对 JEV E基因 5′端设计合成一对特异引物 ,以日本脑炎病毒内蒙古分离株 M73-1 RNA为模板 ,经 RT-PCR扩增 ,获得 366bp的 JEV E基因 c DNA片段 .将此片段重组于质粒 p UC1 9中 ,并转化大肠杆菌DH5α.经 PCR扩增 ,酶切及序列分析 ,结果表明 JEV M73-1 5′端 1 2 6个核苷酸序列与 JEV日本 Nakayama株和 Ja OAr S982株的同源性分别为 96.8%和 96.8% ,氨基酸序列同源性均为 99.2 % .通过对病毒基因组结构分析从分子水平上进一步证实了 1 973年在呼和浩特市流行的脑炎是由 JEV引起的流乙脑炎 .E基因编码 JEV的囊膜糖蛋白 ,是其重要表面抗原 .JEV M73-1 E基因 5′端克隆和部分序列分析对 JEV的分子生物学研究、建立分子生物学诊断技术以及基因工程疫苗的研制均具有重要学术意义和实际价值  相似文献   

9.
将本室克隆的编码复制酶基因3′端约1/2序列及其3′端非编码区的苜蓿花叶病毒中国分离株(AlMV-Ch)RNA23′端cDNA,重组到植物表达载体pROKII中,通过致瘤农杆菌(A-grobacteriumtumefaciens)介导,以叶圆片为转化材料,转化普通烟草,并获得了转基因植株.经卡那霉素抗性选择、PCR检测目的基因证明AlMVRNA23′端基因已整合到转基因烟草的基因组DNA中.转基因植物的攻毒试验表明转基因植株对苜蓿花叶病毒产生高水平的抗性.  相似文献   

10.
从表现花叶、畸形以及坏死症状的加工番茄病株上获得黄瓜花叶病毒分离物,用1对CMV CP基因引物进行RT-PCR,扩增得到了776bp的特异性核苷酸片段。将PCR产物插入到克隆载体PUCm-T中转化大肠杆菌DHSa。经限制酶酶切鉴定,重组克隆中含有与PCR产物大小相同的776bp的插入片段。cDNA全序列分析表明,重组克隆含1个开放阅读框架(ORF),长度为657nt,编码218个氨基酸组成的蛋白。与国内外报道的CMV株系序列比较,所测得的CMV新疆分离物与CMV亚组Ⅰ的核苷酸同源性高达91.0%-98.9%.而与CMV亚组Ⅱ的核苷酸同源性仅在76.4%-78.2%,所测得的CMV新疆分离物与CMV亚组Ⅰ的氨基酸同源性高达94.2%-98.6%;而与CMV亚组Ⅱ的核苷酸同源性仅在79.9%~83.5%.CMV亚组Ⅰ的株系较CMV亚组Ⅱ株系同源关系更密切,所以CMV新疆分离物应该归属于CMV亚组Ⅰ。  相似文献   

11.
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.  相似文献   

12.
何延凌 《科技信息》2008,(4):258-258
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.  相似文献   

13.
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.  相似文献   

14.
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。  相似文献   

15.
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.  相似文献   

16.
AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…  相似文献   

17.
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.  相似文献   

18.
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.  相似文献   

19.
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-  相似文献   

20.
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-  相似文献   

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