用于通路(Gateway)克隆技术的植物表达载体研究进展

通路(Gateway)克隆技术是根据λ噬菌体基因组和大肠杆菌基因组之间的位点专一性重组分子机制开发的一套分子克隆新技术.利用该技术LR反应构建目的基因的表达载体时不需要经过酶切和连接等繁琐而又费时的过程,因此,可以节省很多时间.为了扩大Gateway技术在植物基因工程领域的应用,最近有很多研究机构和研究小组开发了能用于组成型或诱导型表达目的基因、基因沉默、启动子分析、蛋白质亚细胞定位、蛋白质/蛋白质相互作用、多个DNA片段的模块化组装和DNA组片段功能验证等研究用的植物表达载体.该文对这些技术的研究进展进行了综述.     

无核荔枝果实形成差异表达基因cDNA的克隆

采用抑制差减杂交技术(Suppression Subtractive Hybridization,SSH)分离与海南无核荔枝果实形成相关的差异表达基因的cDNA片段,为克隆相关基因提供研究基础。分别以无核荔枝的有核幼果为driver;无核幼果为tester,建立差减cDNA文库。经Reverse Northern Dot-Blot筛选该文库,共获得61个阳性克隆,随机选取17个克隆进行测序,共获得10条非重复序列,对其中较长的7个序列进行同源分析,结果表明:有6个序列在荔枝中为首次报道。     

武铠蛱蝶线粒体基因组全序列测定和分析

【目的】了解闪蛱蝶亚科属间及种间的分子系统进化关系。【方法】采用PCR步移法对武铠蛱蝶 Chitoria ulupi 线粒体基因组全序列进行了测定和分析。基于线粒体基因组13个蛋白质编码基因的核苷酸序列构建了38种鳞翅目昆虫的系统发育树。【结果】分析结果表明,武铠蛱蝶线粒体基因组全长15 279 bp,包括13个蛋白质编码基因、22个tRNA基因、2个rRNA基因和一段长度为391 bp的A+T富含区,基因排列顺序与其他已知近缘种昆虫相同。武铠蛱蝶线粒体基因组中存在很高的A+T含量（79.9％）。13个蛋白质编码基因中,COII以TTG作为起始密码子,COI以CGA作为起始密码子外,其余均为昆虫典型的起始密码子ATN。COII和ND4基因使用了不完全终止密码子T,其余基因均以典型的TAA为终止密码子。在所测得的22个tRNA基因中,除tRNA^Ser(AGN)缺少DHU臂外,其余tRNA均能形成典型的三叶草结构。与其他多数鳞翅目昆虫一样,武铠蛱蝶的A+T富含区中有一段由ATAGAA引导的保守的多聚T结构,长度为21 bp,并散布着一些长短不一的串联重复单元。系统发育树结果显示,总科级别的系统发育关系为：卷蛾总科＋（凤蝶总科＋（螟蛾总科＋(夜蛾总科＋蚕蛾总科＋尺蛾总科)））;在蛱蝶科物种中,武铠蛱蝶与猫蛱蝶Timelaea maculate 亲缘关系最近。【结论】基于分子标记构建的鳞翅目昆虫系统发育关系与传统的形态学分类结果基本一致。     

RNase1 can modulate gut microbiota and metabolome after Aeromonas hydrophila infection in blunt snout bream

Pancreatic ribonuclease (RNase1) of Megalobrama amblycephala exhibits both antimicrobial and digestive activity. The gut microbiome improve the digestion and metabolic capacity and enhance the functioning of the immune system of the host against pathogenic bacteria. In this study, we aimed to assess the protective effect of RNase1 on Aeromonas hydrophila-induced inflammation and intestinal microbial metabolism. Megalobrama amblycephala were randomly divided into three groups: control (injected PBS), infection (A. hydrophila-injected), and treatment group (RNase1 pretreatment 24 h before the A. hydrophila injection). The morphological symptoms were significantly alleviated by RNase1. RNase1 reshaped the perturbed gut microbiota by upregulating Proteobacteria and Vibrio richness and downregulating Firmicutes, Chlamydiae, Bacillus, and Gemmobacter richness. The lysophosphatidylcholine, (±) 17 HETE, D- (+) -cellobiose, and PC (20:5) in the treatment group were restored by RNase 1 protein treatment to the level of the control group. In the treatment group, phospholipid metabolism, fatty acid metabolism, glucose metabolism and lipid metabolism were different from the control and infection groups. The proinflammatory factors concentration in intestinal samples significantly increased after A. hydrophila infection. Our results revealed that RNase1 plays an important role in resistance to pathogen invasion, reducing inflammation, and improving intestinal function, thus inhibiting the occurrence of disease.     

穿心莲中的三个二萜酸盐

从穿心莲（Andrographis paniculata Nees.)水溶性部位分离得到3个新的二萜酸盐：穿心莲酸镁，穿心莲酸二钠和穿心莲酸二钾葡萄糖甙，同时，首次从穿心莲中分得鸟瞟呤核苷，尿嘧啶核苷，6－epiharpagide,procumbide，三色堇黄苷和芹菜素－7－氧－葡萄糖甙。通过波谱解析和化学方法确定了它们的结构。