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转座子标签及其在酿酒酵母基因功能研究中的应用 总被引:2,自引:0,他引:2
转座子标签(transposontagging)技术是研究功能基因的有效的工具之一。介绍了几种在酿酒酵母(Saccharomycescerevisiae)基因功能研究中应用的转座子标签:mTn3标签、miniMu噬菌体标签和Ty转座子标签,阐述了转座子标签的构建原理、应用策略和转座子标签插入位点的鉴定方法。 相似文献
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piggyBac转座子在牛基因组的整合位点及特征分析 总被引:1,自引:0,他引:1
piggyBac(PB)转座子作为一种遗传工具被广泛应用于多个物种的转基因及插入突变研究, 目前PB转座子在牛中的相关研究还较少。为了获得PB转座子在牛基因组中的整合位点, 总结其转座特征, 文章构建了PB[CMV-EGFP]和pcDNA-PBase二元转座系统, 利用细胞核电转技术共转染牛耳组织成纤维细胞, 经G-418筛选, 获得了稳定转染EGFP的转基因细胞系; 提取细胞基因组DNA, 利用基因组步移技术扩增PB转座子5′ Bac区插入位置的DNA序列; 通过与牛基因组序列进行BLAST比对, 得到PB转座子在牛基因组中的插入位点。文章共获得了8个有效的整合位点, 但仅有5个位点定位到染色体1、2、11和X染色体上。序列分析表明:在牛基因组中, PB转座子可特异性的插入到“TTAA”位置, 并整合到基因间的非调控区; 分析整合位点“TTAA”相邻一侧的5个碱基组成, 发现PB转座子5′端倾向于插入到GC(62.5%)碱基富集区。该研究表明, PB转座子可以在牛基因组中发生转座, 获得的整合位点信息为利用PB转座子在牛上开展遗传学研究提供了理论参考。 相似文献
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2.在根瘤菌研究中成功地运用了转座子诱变技术。转座子(Transposon)是一种特殊的DNA短片段,它带有抗药性基因,并具有在DNA复制子之间转座插入的能力,转座的发生并不需要recA基因产物,一些转座子象Tn 5的转座插入位点的分布是相当随机的,但另一些象Tn 10,它的转座插入似乎具有“热点”(Hot spot),转座子插入到一个新位点时,被插入位点原基因的连续性受到阻断,因而该基因的功 相似文献
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piggyBac转座子及其在转基因昆虫中的应用 总被引:1,自引:0,他引:1
piggyBac是一种从粉纹夜蛾Trichoplusiani.中分离到的、具有TTAA插入位点特异性的DNA转座子。piggyBac可在昆虫基因组中准确切离,转化频率较高,并且不受宿主因子的限制,是目前转基因昆虫研究中应用最广的转座子载体。近年来的研究发现,piggyBac类转座子广泛分布于昆虫和其他生物基因组中。文章从piggyBac的结构、转座特性、在转基因昆虫中的应用以及piggyBac类转座子的分布等几个方面综述了piggyBac的研究进展。 相似文献
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转座子挽救法对苜蓿中华根瘤菌与耐盐有关基因的定位 总被引:5,自引:0,他引:5
用含Tn5转座子的质粒pRL1063a诱变苜蓿中华根瘤菌(Sinorhizobium meliloti)042BM,得到盐敏感突变株042BML-2。采用转座子挽救法对Tn5插入位点两边的序列进行克隆与测序,获得了1179bp的转座子插入位点侧翼DNA序列。在GenBank中进行序列同源性和基因定位分析,结果表明:转座子插入在一个功能未知的基因内部,此基因长6270bp。研究证明:该基因与042BM的耐盐性有关,并定名为rtsC。氨基酸疏水性分析表明,在RtsC蛋白的N端有两个跨膜区,该蛋白与细菌趋化性相关蛋白的功能域有同源性。并对RtsC蛋白在苜蓿中华根瘤菌042BM耐盐性中的作用进行了讨论。 相似文献
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铜绿假单胞菌蹭行运动相关基因的研究 总被引:3,自引:0,他引:3
应用Mu转座重组技术研究铜绿假单胞菌 (Pseudomonasaeruginosa)蹭行运动 (Twitchingmotility)的相关基因。通过转座突变、表型筛选 ,得到 8个Twitchingmotility缺陷或减弱的突变子。经过基因克隆、核苷酸测序研究 ,鉴定转座子插入到基因组中的位置。结果表明 ,在其中 4个突变子中 ,转座子分别插入到与IV型菌毛生物合成和功能相关的 3个已知基因中 (其中有两个突变子转座子插入到同一基因的不同位置 ) ,它们是pilV ,pilQ ,algR。另外 4个突变子中 ,有 3个是转座子分别插入到基因pilL基因的前端 ,中部和后端 ,均引起Twitchingmotility功能缺失。另一个突变子中 ,转座子插入到基因PA1 82 1中 ,引起Twitchingmotility功能减弱。PilL和PA1 82 1的编码产物均属于 3 类蛋白质 ,它们的功能是根据其保守的氨基酸基序或基因序列与已知功能基因的相似性推测得出的。但缺乏详细的试验证据。研究结果为pilL控制Twichingmotility提供了有力的证据。并证实基因PA1 82 1与Twitchingmotility有关。将Mu转座重组技术应用到假单胞菌的研究中 ,国内外均未见报道。由于该技术具有随机单点插入的优点 ,克服了传统转座子能在染色体上迁移的缺点。保证了表型的改变与转座子插入位点的基因突变的一一对应关系。为进一步研究铜绿假 相似文献
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转座子在各类真核生物基因组中都占有很高的比例,它们对宿主基因组特别是关联的基因在结构、功能和进化上都起着重要的作用。基于生物信息学分析,本研究选择了水稻基因组中2个被转座子插入的宿主基因,通过PCR扩增和琼脂糖凝胶电泳分析,获得了转座子在稻属16个代表物种94份材料中的插入式样。结果表明,这2个转座子在稻属中的分布式样与插入时间不同,基因三DG-&02926349中的转座子在AA-基因组的物种中全部存在,基因LOC-Os02945130中的转座子则插入稻属AA-基因组的部分物种中,与AA-基因组的物种的系统发育关系相吻合。转座子在宿主基因组中不同的分布与保留式样以及插入后已经固定在不同地理来源的群体中,暗示了它们在物种进化过程中对宿主基因可能存在适应性意义。 相似文献
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转座子是DNA插入因子的一种,是指能在基因组间或组内跳跃的DNA片段。转座子作为插入突变剂或分子标签已被广泛地应用于基因的分离和克隆,且因其独特的性质已成为发现新基因和基因功能分析的有效工具。这使得转座子无论是在单基因水平还是全基因组水平,都成为细菌、酵母和其他微生物研究的有力工具。简单而有效的体外转座反应可以对一些以往难以进行分析的顽固微生物进行转座诱变分析。而建立在转座子基础上的信号标签诱变技术和遗传足迹法的应用则发现了一些新的病原微生物毒力因子,从而可以更好地对这些病原微生物的致病机理进行阐述。这些再次说明转座子是微生物功能基因组研究中的有力工具。本文综述了转座子及其衍生载体介导的一些技术,并讨论其在微生物功能基因组研究中的应用。 相似文献
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Jansen LA 《Bioethics》2006,20(2):105-111
When applying moral principles to concrete cases, we assume a background shared understanding of the boundaries of the persons to whom the principles apply. In most contexts, this assumption is unproblematic. However, in end-of-life contexts, when patients are receiving 'artificial' life-support, judgments about where a person's self begins and ends can become controversial. To illustrate this possibility, this paper presents a case in which a decision must be made whether to deactivate a patient's pacemaker as a means to hasten his death. After discussing some common moral principles that are often applied to resolve ethical problems at the end of life and after explaining why they are of no help here, the paper argues that the correct analysis of this case, and of cases of this sort, turns on considerations that relate to the constitution of the self. These considerations, the paper further argues, sometimes resist resolution. The constitution of the self is fixed in large measure by our concepts and social conventions, and these do not always provide determinate grounds for delimiting the boundaries of the self. 相似文献
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van Veelen M 《Journal of theoretical biology》2005,237(4):412-426
This paper distinguishes two categories of questions that the Price equation can help us answer. The two different types of questions require two different disciplines that are related, but nonetheless move in opposite directions. These disciplines are probability theory on the one hand and statistical inference on the other. In the literature on the Price equation this distinction is not made. As a result of this, questions that require a probability model are regularly approached with statistical tools. In this paper, we examine the possibilities of the Price equation for answering questions of either type. By spending extra attention on mathematical formalities, we avoid the two disciplines to get mixed up. After that, we look at some examples, both from kin selection and from group selection, that show how the inappropriate use of statistical terminology can put us on the wrong track. Statements that are 'derived' with the help of the Price equation are, therefore, in many cases not the answers they seem to be. Going through the derivations in reverse can, however, be helpful as a guide how to build proper (probabilistic) models that do give answers. 相似文献
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Amanda-Jayne F. Carr T. Katherine Tamai Lucy C. Young Veronica Ferrer Marcus P. Dekens David Whitmore 《Chronobiology international》2006,23(1):91-100
Zebrafish are typically used as a model system to study various aspects of developmental biology, largely as a consequence of their ex vivo development, high degree of transparency, and, of course, ability to perform forward genetic mutant screens. More recently, zebrafish have been developed as a model system with which to study circadian clocks. Cell lines generated from early-stage zebrafish embryos contain clocks that are directly light-responsive. We describe recent experiments using single-cell luminescent imaging approaches to study clock function in this novel cell line system. Furthermore, studies examining the process of entrainment to light pulses within this cell population are described in this review, as are experiments examining light-responsiveness of early-stage zebrafish embryos. 相似文献
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Seagrasses are experiencing fragmentation and regression globally; thus, protection and recovery of meadows are a preservation priority. However, conservation actions must consider inherent regional conditions, since certain coastal areas are not suitable for the settlement of extensive meadows. Likewise, small oceanic archipelagos are not always able to fulfil the habitat requirements of seagrass habitats but can harbour small patches that in turn provide unique research opportunities. In this study, we focused on the seagrass Cymodocea nodosa in the archipelago of Madeira (NE Atlantic Ocean). Here we compile historical and contemporary records of this species along with characterization of associated communities (fish and invertebrates). A bionomic map with potentially suitable areas for the establishment and settlement of this species is also included. Lastly, we highlight coastal management and restoration actions and future research directions to preserve this species in Madeira Island. 相似文献
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三峡库区消涨带维管植物区系的初步研究 总被引:13,自引:4,他引:9
三峡库区消涨带是指三峡库区范围内长江进一步充及其支流由于江(河)水水位季节性涨落。导致其沿江(河)两岸土地出现周期性淹没而自然形成的带状区域。该带维管植物区系具有以下特征;植物种类比较丰富,计有维管植物83科,240属,377种,26变种和2变型;地理成分复杂,联系广泛,温带成分占优势;区系组成表现进化性,生活型组成以多年生草本,一年生草本和灌木为主;地方特有成分不多,但中国特有种类比较丰富;带内不同区段物种丰富度有差异。干流多于支流,奉节以东多于奉节以西,带上部多于带中部和带下部;优势现象明显,表征类型丰富。 相似文献
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Carr AJ Tamai TK Young LC Ferrer V Dekens MP Whitmore D 《Chronobiology international》2006,23(1-2):91-100
Zebrafish are typically used as a model system to study various aspects of developmental biology, largely as a consequence of their ex vivo development, high degree of transparency, and, of course, ability to perform forward genetic mutant screens. More recently, zebrafish have been developed as a model system with which to study circadian clocks. Cell lines generated from early-stage zebrafish embryos contain clocks that are directly light-responsive. We describe recent experiments using single-cell luminescent imaging approaches to study clock function in this novel cell line system. Furthermore, studies examining the process of entrainment to light pulses within this cell population are described in this review, as are experiments examining light-responsiveness of early-stage zebrafish embryos. 相似文献
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Amanda‐Jayne F. Carr T. Katherine Tamai Lucy C. Young Veronica Ferrer Marcus P. Dekens 《Chronobiology international》2013,30(1-2):91-100
Zebrafish are typically used as a model system to study various aspects of developmental biology, largely as a consequence of their ex vivo development, high degree of transparency, and, of course, ability to perform forward genetic mutant screens. More recently, zebrafish have been developed as a model system with which to study circadian clocks. Cell lines generated from early‐stage zebrafish embryos contain clocks that are directly light‐responsive. We describe recent experiments using single‐cell luminescent imaging approaches to study clock function in this novel cell line system. Furthermore, studies examining the process of entrainment to light pulses within this cell population are described in this review, as are experiments examining light‐responsiveness of early‐stage zebrafish embryos. 相似文献
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Ligularia speices are widely used in Asian folk medicines for the treatment of various human diseases. Eremophilane‐type sesquiterpenes are abundant and typical secondary metabolites found in this genus. Over 500 eremophilanes reported from members of Ligularia are reviewed in this article together with bioactivity data in an effort to highlight the development in this field. 相似文献