新的端粒酶活性抑制蛋白LPTS-L的制备

LPTS基因是利用定位候选克隆策略克隆的一个新的肝相关候选肿瘤抑制基因。LPTS基因编码一个全长为328氨基酸的蛋白质(LPTS-L),该蛋白具有抑制细胞端粒酶活性的功能。为了进一步研究LPTS-L蛋白的结构与功能,利用DNA重组技术,将LPTS-L的cDNA克隆到表达载体pET-24a中构建重组克隆pET-24-LPTS,并在大肠杆菌BL-21中进行融合表达,获得可溶形式的LPTS-L融合蛋白。采用Ni Sepharose4B柱亲和层析,可以获得纯度较高的蛋白,但不适合大量制备。通过设计引物去掉了pET-24a载体上的6×His tag将LPTS-L基因进行了非融合表达,然后采用磷酸纤维素P11阳离子交换层析纯化LPTS-L蛋白,纯度可达到55%。再经Sephadex G-100凝胶过滤,LPTS-L蛋白的纯度可达到80%。Western blot实验显示经纯化后的LPTS-L蛋白可与兔抗GST-LPTS-L的多抗发生特异性结合。采用TRAP法测定蛋白质活性,结果显示纯化得到的LPTS-L蛋白可抑制端粒酶的活性,与采用Ni Sepharose4B纯化获得的LPTS-L融合蛋白比较,其抑制效率基本一致。因此,所建立的技术可以有效地制备LPTS-L蛋白。     

甘肃蝴蝶名录

基于近几年的野外工作,结合前人对甘肃省蝶类的研究,对甘肃省蝴蝶名录进行了整理,并对各分类阶元进行了统计。截至2013年,甘肃省共记录蝶类12科210属614种,科、属、种的数量分别占中国总数量的100%、57.22%和28.52%。按照中国动物地理区划,古北种285种,东洋种233种,广布种95种。甘肃省境内分布有各类珍稀濒危蝶类37种,其中国家Ⅱ级重点保护野生动物2种(凤蝶科的三尾凤蝶和绢蝶科的阿波罗绢蝶),列入《国家保护的有益的或者有重要经济、科学研究价值的野生动物名录》的有11属33种,列入《濒危野生动植物种国际贸易公约》的有4种,列入《世界濒危物种红色名录》的有5种。     

HAL1 mediate salt adaptation in Arabidopsis thaliana

INTRODUCTIONSalinity is a major environmental stress that isa substantial constraint to crop production both fordry land and irrigated agriculture. The detrimental impact of this stress is perpetuated and exacerbated by management practices used to facilitatehigh-output crop production. To overcome theselimitations and improve production efficiency in theface of a burgeoning world population, more salt tolerant crops must be developed. In contrast with traditional breeding, the direct ill…     

入侵植物加拿大一枝黄花与乡土植物芦苇的相互化感作用

以加拿大一枝黄花和芦苇的共优群落和各自单优群落的种子为化感受体,分析其分别在加拿大一枝黄花和芦苇5个浓度梯度(12.5、25、50、100和200mg·mL-1)浸提液处理下的发芽率和芽长差异,研究2种植物的相互化感作用规律.结果表明:2种植物在共优群落中的千粒重和蒸馏水培养下的发芽率均比单优群落大.加拿大一枝黄花的浸提液对自身种子发芽率在单优和共优群落中均具有在低浓度下(12.5、25 mg·mL-1)轻微促进、中高浓度下(50、100和200mg·mL-1)强烈抑制的作用,其中对共优群落种子的抑制作用尤为显著;而芦苇浸提液对加拿大一枝黄花种子的影响无明显规律.加拿大一枝黄花的种子芽长在单优和共优群落中均随自身浸提液浓度的升高趋于减小,随芦苇浸提液浓度的升高呈波动减小趋势.在芦苇和加拿大一枝黄花浸提液处理下,芦苇种子发芽率在单优群落中均显著大于共优群落(P＜0.05),但2种浸提液处理间无显著差异.     

用RACE技术扩增并克隆牛BMP4基因3'端序列

RACE技术是一项扩增基因末端序列的新技术。该研究从牛BMP4基因出发,以牛软骨的RNA为模板,按照不同物种BMP4基因的相似性设计特异引物,运用PCR和RACE技术扩增并获得了特异片段,该片段经PCR、酶切和测序验证,证实所克隆序列为牛BMP4的3′端序列,包含有1170bp组成的开放读码框(ORF),编码389个氨基酸,3′非编码区121bp个核苷酸和poly(A)15。同源性分析结果表明,牛BMP4 cDNA最大开放读码框所推测的氨基酸序列与已知人、小鼠、大鼠、狗、羊和鸡等真核生物BMP4氨基酸序列进行比较,分别有94.5%、93.1%、91.9%、87.4%、94.2%、79%的同源性。这为克隆其他物种的BMP4基因提供了依据,同时牛骨形态发生蛋白的测序为我们更好的理解牛的生骨机理提供帮助。     

丘陵地区地形梯度上植被格局的分异研究概述

植物群落的本质特征之一是群落中的植物和环境之间存在一定的相互关系。湿润的丘陵地区是由水侵蚀而形成的包含各种干扰频率的生境复合体,作为中尺度的地形单位,可以通过侵蚀前线划分为上部坡面和下部坡面两个小尺度的地形单位,而上部坡面可以进一步划分为顶坡、上部边坡、谷头凹地等微地形单元,下部坡面可以进一步划分为下部边坡、麓坡、泛滥性阶地及谷床等微地形单元。上部坡面发育的是气候顶极群落,沿顶坡向谷头凹地,群落发生逐渐、连续的变化,下部坡面发育的为地形群落,其物种组成、结构以及其它生态特征与上部坡面具有显著的差异,而其微地形单元之间植被的变化不明显。干扰作用是不同地形植被分异的控制因子,也是地形植被维持和更新的关键因子。下部坡面以相对积极的土壤侵蚀、滑坡和崩塌等过程为特征,其植被更新依赖于频繁的地面干扰,而上部坡面长期稳定,其植被更新依赖于林窗动态。地形是影响植被格局的最重要的也是最基本的生境因子,其引起的生境生态位分化为物种的共存提供了条件,导致了小尺度空间内高生物多样性的形成和维持。     

Horizontal transmission, vertical inactivation, and stochastic loss of mariner-like transposable elements

Horizontal transmission has been well documented as a major mechanism for the dissemination of mariner-like elements (MLEs) among species. Less well understood are mechanisms that limit vertical transmission of MLEs resulting in the "spotty" or discontinuous distribution observed in closely related species. In this article we present evidence that the genome of the common ancestor of the melanogaster species subgroup of Drosophila contained an MLE related to the mellifera (honey bee) subfamily. Horizontal transmission, approximately 3-10 MYA, is strongly suggested by the observation that the sequence of the MLE in Drosophila erecta is 97% identical in nucleotide sequence with that of an MLE in the cat flea, Ctenocephalides felis. The D. erecta MLE has a spotty distribution among species in the melanogaster subgroup. The element has a high copy number in D. erecta and D. orena, a moderate copy number in D. teissieri and D. yakuba, and was apparently lost ("stochastic loss") in the lineage leading to D. melanogaster, D. simulans, D. mauritiana, and D. sechellia. In D. erecta, most copies are concentrated in the heterochromatin. Two copies from D. erecta, denoted De12 and De19, were cloned and sequenced, and they appear to be nonfunctional ("vertical inactivation"). It therefore appears that the predominant mode of MLE evolution is vertical inactivation and stochastic loss balanced against occasional reinvasion of lineages by horizontal transmission.      

A phylogenetic approach to the identification of phosphoglucomutase genes

The expanding molecular database provides unparalleled opportunities for characterizing genes and for studying groups of related genes. We use sequences drawn from the database to construct an evolutionary framework for examining the important glycolytic enzyme phosphoglucomutase (PGM). Phosphoglucomutase plays a pivotal role in the synthesis and utilization of glycogen and is present in all organisms. In humans, there are three well-described isozymes, PGMI, PGM2, and PGM3. PGM1 was cloned 5 years ago; however, repeated attempts using both immunological approaches and molecular probes designed from PGM1 have failed to isolate either PGM2 or PGM3. Using a phylogenetic strategy, we first identified 47 highly divergent prokaryotic and eukaryotic PGM-like sequences from the database. Although overall amino acid identity often fell below 20%, the relative order, position, and sequence of three structural motifs, the active site and the magnesium-- and sugar-binding sites, were conserved in all 47 sequences. The phylogenetic history of these sequences was complex and marked by duplications and translocations; two instances of transkingdom horizontal gene transfer were identified. Nonetheless, the sequences fell within six well-defined evolutionary lineages, three of which contained only prokaryotes. Of the two prokaryotic/eukaryotic lineages, one contained bacterial, yeast, slimemold, invertebrate, and vertebrate homologs to human PGM1 and the second contained likely homologs to human PGM2. Indeed, an amino acid sequence, derived from a partial human cDNA, that fell within the second cross-kingdom lineage bears several characteristics expected for PGM2. A third lineage may contain homologs to human PGM3. On a general level, our phylogenetic-based approach shows promise for the further utilization of the extensive molecular database.      

Short chain oligogalacturonides induce ethylene production and expression of the gene encoding aminocyclopropane 1-carboxylic acid oxidase in tomato plants

Oligogalacturonic acids (OGAs), derived from plant cell wall pectin, have been implicated in a number of signal transduction pathways involved in growth, development and defense responses of higher plants. This study investigates the size range of OGAs capable of inducing ethylene synthesis in tomato plants, and demonstrates that in contrast with many other effects, only short chain OGAs are active. Oligomers across a range of DP from 2-15 were separated and purified to homogeneity by QAE-Sephadex anion exchange chromatography using a novel elution system. The OGAs were applied to tomato plants and assayed for their ability to induce ethylene gas release and changes in steady state levels of mRNA encoding the ethylene forming enzyme aminocyclopropane-1-carboxylic acid oxidase (ACO). The study demonstrated that only OGAs in the size range of DP4-6 were active both in eliciting ACO expression and in the production of ethylene.      

Release of Membrane-Bound Trails by Trypanosoma cruzi Amastigotes onto Modified Surfaces and Mammalian Cells

ABSTRACT. Upon incubation at 37° C onto glass coverslips coated with Concanavalin A, poly-L-lysine, or a monoclonal antibody (1D9) directed to the parasite major surface glycoprotein Ssp-4, extracellular Trypanosoma cruzi amastigotes release trails of material barely visible by light microscopy. This release is not associated with parasite movements. Immunolabeling studies confirmed that the material is derived from the parasite's membrane since thin section through samples labeled with 1D9 revealed that the trails are membrane-bound structures. Scanning electron microscopy showed that the ∼0.1-μm thick trails of material emerging from the amastigotes can be uniform or beaded, indicating a tendency to vesiculation. The trails are preferentially released from the flagellar pocket region and/or at the opposite posterior end of the parasite body, and seem to be devoid of microtubules. The release is time and temperature-dependent and fixed parasites do not form trails. All attempts to inhibit trail release using drugs (antimycin A, sodium azide, cytochalasin D, nocodazole, genistein, staurosporine, EGTA) failed. The observation of trails associated with intracellular parasites and amastigotes invading Vero cells suggests that this is probably a physiological process.