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1.
HCG抗原决定簇与乙肝病毒核心抗原的融合表达   总被引:3,自引:0,他引:3  
利用PCR方法获得编码人绒毛膜促性腺激素β链羧端37肽基因片段(HCG-β-37-CTP),并将其分别和乙肝核心抗原的氨端(第1位)、羧端(第454位)、中间(第75~83位)以及中间和羧端同时融合,构建T4个重组融合表达克隆:pCn-HCG、pCc-HCG、pCm-HCG和pC-HCG2,在大肠杆菌中实现了表达。对表达产物中的乙肝核心抗原和HCG抗原的抗原性和表达水平、融合蛋白的颗粒性及其免疫原性进行了分析。其中pCm-HCG和pCc-HCG能形成颗粒。用pCm-HCG免疫小鼠能产生高滴度的抗HCG抗体,说明核心抗原的中间部位是合适的融合位点。  相似文献   

2.
哺乳类细胞中周期蛋白依赖激酶抑制因子   总被引:5,自引:0,他引:5  
近两年,人们相继发现了一组能结合性抑制周期蛋白依赖激酶(CDKs)活性的蛋白因子──CDK抑制因子(CKIs)。它们分别是:p21、p16、p15、p27和CDIl。p21和p27有一定同源性,能抑制多种CDKs的活性;p16和p15则同源性更高,能特异地与CDK4、CDK6结合;CD11的结合特异性还有待进一步的研究。p21的表达受p53的正调控;TGF-β则上调p15的表达以及p27的抑制活性。以上表明CKls不仅是CDKs活性的调控者,而且还是抑癌因子与细胞周期调控之间的直接联系者。  相似文献   

3.
大鼠皂素蜕膜心肌肌钙蛋白Ca^2+敏感性的实验研究   总被引:3,自引:0,他引:3  
采用500μg/ml皂素溶液浸浴大鼠右室乳头肌30min制备蜕膜心肌纤维标本。经撤除MgATP法检验,肌膜通透性明显增高。用含不同浓度Ca ̄(2+)(以Ca ̄(2+)浓度负对数pCa表示)的激活液进行顺序激活,记录Ca ̄(2+)激活张力,其张力-pCa关系曲线描述了肌钙蛋白(TN)对Ca ̄(2+)的亲和特性;曲线中位值pCa_(50)(即产生50%最大张力所对应的pCa)是反映TNCa ̄(2+)敏感性的定量指标。本实验观察到大鼠右室乳头肌张力-pCa关系曲线呈S形,测得pCa_(50)为5.727±0.086(n=16);撤除激活液中磷酸肌酸和磷酸肌酸激酶,张力-pCa曲线左移位,pCa_(50)增高至6.194±0.121(P<0.01)。  相似文献   

4.
利用PCR技术和DNA体外重组方法,把作为导向效应细胞到靶部位的单核细胞趋化激活因子(MCAF)和粒细胞巨噬细胞集落刺激因子(GM-CSF)进行基因融合,置于pBV220载体的λPRPL串联启动子下游,构建了SD序列与ATG之间含有不同核苷酸组成的重组质粒pMG01、pMG02和pMG03。pMG01、pMG02和pMG03的翻译起始区都不存在稳定的二级结构,但DH5α(pMG02、DH5α(pMG03)的表达水平远远高于DH5α(pMG01),DH5α(PMG01)几乎没有表达。表达产物经Westernblot检测表明,它能分别与MCAF和GM-CSF抗体发生特异反应。生物学活性测定表明,表达产物具有明显的单核细胞趋化活性和维持hGM-CSF依赖的TF1细胞生长的特性,说明MCAF和GM-CSF的生物学功能是相容的.  相似文献   

5.
利用从真养产碱杆菌(Alcaligeneseutrophus)H16染色体DNA中克隆到的催化聚β羟基丁酸酯(polyβhydroxybutyrate,PHB)生物合成的两个关键酶基因:依赖NADPH的乙酰乙酰CoA还原酶基因(phbB)以及PHB合酶基因(phbC),同时利用大肠杆菌高效表达载体pKK2233,构建了嵌合有phbB和phbC基因的表达载体pKCB,转入大肠杆菌JM109,通过显微镜观察及气相色谱分析检测PHB的合成。在确证克隆基因正确的基础上构建了马铃薯块茎特异性表达载体pPSAGB(含phbB基因)、pBIBGC(含phbC基因)和pPSAGCB(含phbB和phbC双基因),转化5个马铃薯品种,经检测获得20个转基因阳性株系。  相似文献   

6.
将编码丙型肝炎病毒(HCV)E2蛋白417~750位氨基酸的DNA片段 克隆到真核表达载体pcDNA 3.1(-)中的CMV IE启动子下游,构建成HCV E2重组真核表达质粒 pcE2。ELISA法检测pcE2 DNA免疫兔血清中的E2抗体变化和维持规律,结果显示免疫20d已有 抗体产生,30d后开始进入高峰,40d时达到最高值,至第90d抗体水平保持平稳,抗体滴度 达到1∶1600左右。流式细胞计数仪(FACS)检测pcE2 DNA免疫鼠CD4+、CD8+T淋巴细胞变 化情况,与注射空载体pCDNA3.1(-)的阴性鼠相比,CD4+淋巴细胞水平略有上升,CD8+ 细胞水平有较大升高,增幅达35.46%。免疫组化检测结果显示注射pcE2的小鼠组织中有明显 的阳性着色,而注射pcDNA3.1(-)的对照组小鼠免疫组化结果为阴性。以上结果表明:pcE2 在实验动物内表达出的HCV E2蛋白可以引起免疫动物的体液免疫应答和细胞免疫应答,尤其 是MHC-1限制性杀伤性CD8+T淋巴细胞水平的提高对清除 病毒是十分有利的,因此HCV E2 DNA免疫有可能成为预防和治疗HCV感染的一条新途径。  相似文献   

7.
血管平滑肌细胞增殖与Cdk抑制蛋白p27的表达   总被引:5,自引:1,他引:4  
Yuan Y  Xu DL  Liu YL  Jia MY 《生理学报》1999,51(3):285-290
p27蛋白是细胞周期素依赖性激酶(Cdk)抑制蛋白家族中的一种,主要对外部促进或抑制细胞增殖的信号起反应。本研究应用流式细胞仪(FCM)双标记的方法观察血管紧张素Ⅱ(AngⅡ)、血管加压素(AVP)和血小板源生长因子(PDGF)对血管平滑肌细胞(VSMCs)细胞周期百分比和p27蛋白表达量的影响。静止状态培养的VSMCs加入AngⅡ,AVP,PDGFBB后,在不同时间收集细胞,用碘化丙啶(PI)标记细胞DNA,以确定细胞所处的周期。用p27蛋白的单抗和标记了FITC的二抗标记细胞,通过流式细胞仪测定被激发出的荧光量来确定细胞p27蛋白表达的相对量。结果显示,AngⅡ刺激VSMCs增生,其蛋白含量增加了436%(P<001),但不抑制p27蛋白的表达;AVP可轻度抑制p27的表达,有轻度促进VSMCs增殖和增生的作用(P<005);PDGF明显抑制p27的表达,引起细胞增殖。本研究结果提示,p27蛋白抑制VSMCs通过G1期进入S期,是抑制VSMCs增殖的重要调节因子。  相似文献   

8.
人I型胶原基因第一内含子调节转录的研究   总被引:3,自引:0,他引:3  
人I型胶原α1(I)链(COLIA1)基因内含子序列在不同细胞内有不同的转录调节活性,报道了含人COLIA1基因内含子I不同区段(+544~+855和+820~+1093)的重组质粒pSCEP-CAT和pSCIP-CAT的构建并转染人胚肌腱成纤维细胞和Tca8113舌癌细胞,地高辛标记抗CAT-ELISA检测结果显示:pSCEP-CAT在两种细胞均获表达;pSCIP-CAT在人成纤维细胞未表达,但  相似文献   

9.
宽阔水自然保护区的虫草及其相关真菌Ⅱ.   总被引:2,自引:0,他引:2  
采自贵州省宽阔水自然保护区的另6种虫草,它们是娄山虫新种(CordycepsloushanensisLiang&Liusp.nov)。绿核虫草新种(CordycepsaeruginosclerotaLiang&Liusp.Shimazu),拟暗绿虫草(CordycepspseudoatrovirensKob&.Shim),布氏虫草(CordycepsbrongniartiiShimazu)金针虫虫  相似文献   

10.
用三亲水交配方法分别将载有褐球固氮菌(Azotobacterchroococcum)呈组成型表达的nifAC的质粒pCK5和肺炎克氏杆菌(Klebsiellapneumoniae)含有nifA^C和nifA-ntrC基因的质粒pCK3,pSZ36和pSZ23-CA导入根癌土壤杆菌(Agrobacteriumtunefaciens)C58/pGV3850所得转移接合子的生长速率和野生型相似。在10m  相似文献   

11.
湿地克隆植物的繁殖对策与生态适应性   总被引:27,自引:0,他引:27  
湿地是地球上独特的生态系统,它是水陆交替的过渡带,而湿地植物为了适应这种特殊的环境,必须有其相应的生长格局(growthpattern)和繁殖对策(reproductivestrategy)。有些湿地植物具有密丛型的生态特点,这是对过湿的土壤和氧气供给不足环境的一种适应方式。随着有机质的不断积累,每年从分蘖节向上长出新枝,向下生长不定根,其分蘖节不断上移,以便从地表面上获得氧气,并使植物体免遭埋没,这样年复一年的生长,在地表面上形成了草丘。生殖是生物繁衍后代延续种族最基本的行为和过程,它不仅是…  相似文献   

12.
Ligularia virgaurea is a perennial herb that is widely distributed in the alpine meadow on the eastern Qinghai-Tibet plateau.We investigated the patterns of growth and reproduction of L.virgaurea under two contrasting levels of light conditions for two continuous growing seasons.Our results showed that the light affects on the maximum relative growth rate,the shoot weight ratio and the root weight ratio differed between the two growing seasons.L.virgaurea reproduced initially through rhizome in the second growing season,rather than sexual reproduction.The proportion of genets with clonal reproduction decreased under shaded conditions.A minimum genet size should be attained for clonal reproduction to begin under the shaded conditions.There was a positive linear relationship between clonal reproduction and genet size.Light level affected the allocation of total biomass to clonal structures,with less allocation under the full natural irradiance than under the shaded conditions.There seemed to be a trade-off between vegetative growth and clonal reproduction under the full natural irradiance,in terms of smaller relative growth rates of genets with clonal reproduction than those without clonal reproduction.L.virgaurea emphasized clonal reproduction under the full natural irradiance,while the plant emphasized vegetative growth under the shaded conditions.  相似文献   

13.
That trade-offs result from the allocation of limited resources is a central concept of life history evolution. We quantified trade-offs between sexual and clonal reproduction in the aquatic plant, Butomus umbellatus, by experimentally manipulating sexual investment in two distinct nutrient environments. Increasing seed production caused a significant but nonlinear trade-off. Pollinating half of all flowers strongly reduced clonal bulbil production, but pollinating the remaining flowers did not cause any further trade-off. Trade-offs were not stronger under low nutrient conditions that clearly limited plant growth. Experimentally induced trade-offs were not reflected in negative phenotypic correlations between sexual and clonal allocation among plants within eight populations grown in a uniform greenhouse environment. Diminishing effects of increased sexual allocation plus a lack of accord between experimental manipulations and phenotypic correlations suggest that trade-offs between sexual and clonal reproduction are unlikely to constrain the evolution of reproductive strategy in this species.  相似文献   

14.
Abstract. Question: How do the relative frequencies of plant traits (clonality, growth form, seed weight, diaspore morphology) vary during the life cycle and how does this affect regeneration? Location: Alpine meadow and heath communities at Kilpisjärvi, sub‐Arctic Finland. Methods: Control plots and three treatments were used to measure relative species abundances for five life cycle stages: standing vegetation, seed rain, seed bank and seedlings emerging in gaps and in closed vegetation. Results: The relative frequencies of plant traits varied between the life cycle stages. The meadows were dominated by weakly clonal herbs, small or intermediate seeds and unappendaged diaspores, while the heaths were dominated by clonal dwarf shrubs, small seeds and fleshy fruits. In the meadows, species with small seeds dominated during the seed rain and in the seedling stage in gaps, while species with intermediate seeds dominated the seed bank and the seedling stage in closed vegetation. Species with unappendaged diaspores dominated throughout the life cycle. In the heaths, seed bank and seedling stage were practically absent. Conclusions: The observed differences in plant trait spectra between life cycle stages indicate that important environmental factors differ among the stages. Small seeds are advantageous for dispersal, whereas intermediate seeds have a greater probability of germinating and establishing in closed vegetation. Appendages facilitate dispersal, whereas unappendaged diaspores favour seed burial. Although the plant growth form spectrum largely reflects environmental constraints during the regeneration cycle, information on seed weight and diaspore morphology improves our knowledge of the relative importance of morphological adaptations of sexual structures in different stages during the life cycle.  相似文献   

15.
Many aquatic and riparian plant species are characterized by the ability to reproduce both sexually and asexually. Yet, little is known about how spatial variation in sexual and asexual reproduction affects the genotypic diversity within populations of aquatic and riparian plants. We used six polymorphic microsatellites to examine the genetic diversity within and differentiation among 17 populations (606 individuals) of Sparganium emersum, in two Dutch-German rivers. Our study revealed a striking difference between rivers in the mode of reproduction (sexual vs. asexual) within S. emersum populations. The mode of reproduction was strongly related to locally reigning hydrodynamic conditions. Sexually reproducing populations exhibited a greater number of multilocus genotypes compared to asexual populations. The regional population structure suggested higher levels of gene flow among sexually reproducing populations compared to clonal populations. Gene flow was mainly mediated via hydrochoric dispersal of generative propagules (seeds), impeding genetic differentiation among populations even over river distances up to 50 km. Although evidence for hydrochoric dispersal of vegetative propagules (clonal plant fragments) was found, this mechanism appeared to be relatively less important. Bayesian-based assignment procedures revealed a number of immigrants, originating from outside our study area, suggesting intercatchment plant dispersal, possibly the result of waterfowl-mediated seed dispersal. This study demonstrates how variation in local environmental conditions in river systems, resulting in shifting balances of sexual vs. asexual reproduction within populations, will affect the genotypic diversity within populations. This study furthermore cautions against generalizations about dispersal of riparian plant species in river systems.  相似文献   

16.
克隆植物中国沙棘生长对外源植物激素的响应   总被引:1,自引:0,他引:1  
关于植物克隆生长调节问题,目前集中于外在机制的研究。为了探讨中国沙棘克隆生长调节的内在机制,采用3×3回归设计进行田间试验,期望了解不同生长性状对IAA和CTK用量及其配比的响应规律。结果表明:(1)生长性状对激素用量的响应规律呈典型的钟形曲面模式,即各生长指标均存在一个产量峰值,峰值以前生长指标随IAA、CTK用量的增大而提高,峰值以后生长指标随IAA、CTK用量的增大而下降。(2)不同生长指标对激素用量及其配比的响应规律具有一定差异,较高的IAA比例有利于促进树高生长,较高的CTK比例有利于促进地径和冠幅生长,而几乎相等的IAA和CTK用量有利于种群生物量积累。(3)在激素用量适宜的情况下,中国沙棘生长潜力得到充分发挥,形成高大的个体,较多的子株,有利于提高种群对生境资源的占据和利用,并提高排斥其他植物种类入侵的能力;当激素用量过高或过低时,中国沙棘以降低生长量为代价,形成矮小的个体,减少子株数量,有利于削弱个体之间的竞争。这一结果为了解中国沙棘克隆生长内在调节机制提供了线索。(4)根据不同生长指标的激素效应方程,求出了相应的IAA和CTK的最佳用量和最佳配比,以及合理施激素区域和最低成本线。(5)克隆子株数量增幅与地径和冠幅生长量增幅呈极显著正相关、与种群生物量增幅呈显著正相关,即适宜的IAA和CTK用量既可加速个体生长、也能促进克隆子株的产生。  相似文献   

17.
刘凤红  刘建  董鸣 《生态学报》2005,25(12):3415-3419
克隆植物根据其构型可以分为游击型和密集型。游击型克隆植物的间隔子长,分株在水平空间的扩展范围大,可以利用更大空间范围内的资源,其通过克隆生理整合作用发生的非局部反应的能力强。由此可以得出的推论之一是,小生境斑块的环境发生变化,生长于其中的密集型克隆植物的反应可能会更灵敏。这种反应可能会体现在生物量以及配置格局的变化上。以毛乌素地区沙生半灌木群落中两种优势克隆植物羊柴(H edy sarum laeve)和油蒿(A rtem isia ord osica)为研究对象,前者是典型的游击型克隆植物,后者是密集型克隆植物。采取野外调查的方式,观测在不同植被盖度的小生境斑块内二者地上生物量分配格局的变化情况,并结合二者的克隆构型和生活史特征试图探讨产生这种格局的原因。结果表明:羊柴的地上各部分生物量对植被盖度变化的响应不如油蒿敏感。这或者是因为羊柴的游击型克隆构型决定其可以跨越小尺度斑块实现克隆生理整合,可以利用不同小生境斑块的资源导致的。油蒿只能利用小生境斑块内的资源,当小生境斑块的条件改变,其生物量以及配置方式也随之发生相应的变化。在繁殖方式上,羊柴的有性繁殖结构以及有性繁殖投资显著小于油蒿。在资源有限的条件下,对一种繁殖方式的投资常常会削弱另一种繁殖方式。羊柴主要依靠克隆繁殖,这或者符合并支持配置理论的观点。  相似文献   

18.
Nutrient addition to grasslands consistently causes species richness declines and productivity increases. Competition, particularly for light, is often assumed to produce this result. Using a long-term dataset from North American herbaceous plant communities, we tested whether height and clonal growth form together predict responses to fertilization because neither trait alone predicted species loss in a previous analysis. Species with a tall-runner growth form commonly increased in relative abundance in response to added nitrogen, while short species and those with a tall-clumped clonal growth form often decreased. The ability to increase in size via vegetative spread across space, while simultaneously occupying the canopy, conferred competitive advantage, although typically only the abundance of a single species within each height-clonal growth form significantly responded to fertilization in each experiment. Classifying species on the basis of two traits (height and clonal growth form) increases our ability to predict species responses to fertilization compared to either trait alone in predominantly herbaceous plant communities. Electronic supplementary material The online version of this article (doi:10.1007/s00442-012-2264-5) contains supplementary material, which is available to authorized users.  相似文献   

19.
It has been suggested that clonality provides reproductive assurance in cross-fertilizing species subject to pollen limitation, relieving one of the main selective pressures favoring the evolution of self-fertilization. According to this hypothesis, cross-fertilizing species subject to pollen limitation should often be clonal. Here, we investigated the association between clonality and a genetic mechanism enforcing outcrossing, self-incompatibility, in Solanum (Solanaceae). We collected self-incompatibility and clonality information on 87 species, and looked for an association between these two traits. To account for the contribution of shared evolutionary history to this association, we incorporated phylogenetic information from chloroplast (NADH dehydrogenase subunit F) sequence data. We found that self-incompatibility is strongly associated with clonal reproduction: all self-incompatible species reproduce clonally, while the absence of clonality is widespread among self-compatible taxa. The observed correlation persists after taking into account shared phylogenetic history, assumptions about the evolutionary history of self-incompatibility, uncertainty associated with phylogeny estimation, and associations with life history (annual/perennial). Our results are consistent with the hypothesis that clonality provides reproductive assurance, and suggest that the consequences of clonal growth in the evolution of plant reproductive strategies may be more significant than previously thought.  相似文献   

20.
Most plants can reproduce both sexually and asexually (or vegetatively),and the balance between the two reproductive modes may vary widely between and within species.Extensive clonal growth may affect the evolution of life history traits in many ways.First,in some clonal species,sexual reproduction and sex ratio vary largely among populations.Variation in sexual reproduction may strongly affect plant's adaptation to local environments and the evolution of the geographic range.Second,clonal growth can increase floral display,and thus pollinator attraction,while it may impose serious constraints and evolutionary challenges on plants through geitonogamy that may strongly influence pollen dispersal.Geitonogamous pollination can bring a cost to plant fitness through both female and male functions.Some co-evolutionary interactions,therefore,may exist between the spatial structure and the mating behavior of clonal plants.Finally,a trade-off may exist between sexual reproduction and clonal growth.Resource allocation to the two reproductive modes may depend on environmental conditions,competitive dominance,life span,and genetic factors.If different reproductive modes represent adaptive strategies for plants in different environments,we expect that most of the resources should be allocated to sexual reproduction in habitats with fluctuating environmental conditions and strong competition,while clonal growth should be dominant in stable habitats.Yet we know little about the consequence of natural selection on the two reproductive modes and factors which control the balance of the two reproductive modes.Future studies should investigate the reproductive strategies of clonal plants simultaneously from both sexual and asexual perspectives.  相似文献   

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