单链抗体展示系统研究进展

单链抗体(single chain antibody fragment,scFv)是由抗体重链可变区(variable region of heavy chain,VH)和轻链可变区(variable region of light chain,VL)通过柔性短肽连接组成的小分子,是具有完整抗原结合活性的最小功能片段,包含抗体识别及抗原结合部位。相比于其他抗体,scFv具有分子量小、穿透性强、免疫原性弱、易构建表达等优点。目前,scFv最常用的展示系统主要有噬菌体展示系统、核糖体展示系统、mRNA展示系统、酵母细胞表面展示系统和哺乳动物细胞展示系统等。近年来,随着scFv在医学、生物学、食品安全学等领域的发展,使得其在生物合成和应用研究方面备受关注。本文对近年来scFv展示系统的研究进展作一综述,以期为scFv的筛选及应用提供理论基础。     

钾对兰州百合鳞茎中多酚类物质积累代谢及其抗氧化活性的影响

为探究施钾对兰州百合鳞茎中多酚类物质的积累、抗氧化能力及差异代谢物的影响,该研究以兰州百合鳞茎为试材,通过固定氮素(N)和磷素(P)用量,设置不同钾(K)浓度处理,即K_(0)(不施肥)、K_(1)(447.6 mg·L^(-1))、K_(2)(671.4 mg·L^(-1))、K_(3)(895.2 mg·L^(-1)),采用福林-肖卡法、溴甲酚绿比色法、香草醛比色法、DPPH法、铜离子还原能力(CUPRAC)法测定不同K浓度处理下兰州百合鳞茎中多酚类物质含量及其抗氧化活性,并采用LC-MS法分析多酚类物质的差异代谢物,并进行差异代谢物筛选,功能注释及富集分析,为兰州百合的优质栽培提供理论依据。结果表明:(1)不同K浓度处理下兰州百合鳞茎中多酚类物质的含量及其抗氧化活性存在显著差异(P<0.05),与K_(0)相比,K_(1)、K_(2)、K_(3)均能促进鳞茎多酚类物质的积累及其抗氧化能力的提高,其中以K_(2)(671.4 mg·L^(-1))效果最佳。(2)相关性分析表明,兰州百合鳞茎多酚类物质含量与抗氧化活性指标呈极显著(P<0.01)相关关系,相关系数为0.451~0.959。(3)K_(0)、K_(2)浓度处理下兰州百合鳞茎中存在89种多酚类及相关化合物,其中52种相对含量显著上调,37种相对含量显著下调,且显著富集到黄酮类及苯丙类化合物生物合成的通路上。研究认为,兰州百合的最佳施钾量(671.4 mg·L^(-1))能有效促进鳞茎中多酚类物质的积累并提高其抗氧化能力。     

褪黑素调节烟草丁香假单胞菌抗性的功能研究

为分析褪黑素(N-乙酰-5-甲氧基色胺)在植物先天免疫中的功能及调控机理,研究以病原菌丁香假单胞杆菌(Pseudomonas syringae pv.tomato DC3000,Pst DC3000)—烟草互作系统为模型,检测了病原菌侵染对烟草褪黑素相关基因表达的影响,并探讨了褪黑素对植物叶片病原菌生长以及气孔开度和活性氧自由基(reactive oxygen species,ROS)含量的影响以及调控机理。结果表明:(1)Pst DC3000处理提高了烟草褪黑素合成(NtSNAT1)和受体(NtPMTR1)基因表达,且外源褪黑素处理降低了叶片中的病原菌含量。(2)与野生型植物相比,过表达大豆GmSNAT1基因显著提高了转基因烟草中内源褪黑素含量和NtPMTR1的表达,且转基因烟草叶片中的Pst DC3000菌落数显著下降。(3)外源褪黑素和细菌鞭毛蛋白多肽flg22处理诱导了野生型和转基因烟草保卫细胞中ROS产生和气孔关闭,且转基因植物对褪黑素和flg22诱导的气孔关闭和ROS产生比野生型烟草更加敏感。综上所述,研究表明褪黑素可能通过受体NtPMTR1介导的信号途径促进保卫细胞ROS产生,诱导气孔关闭,从而降低病原菌Pst DC3000的入侵。     

西藏色季拉山暗针叶林苔藓多样性及林窗干扰的影响

为了解西藏色季拉山暗针叶林苔藓多样性及林窗干扰的影响,在过去工作积累的基础上研究了色季拉山西坡5块样地内不同林内环境(林窗、林缘和林下)地面、腐木和树附生苔藓生物量特征。结果表明, 地面生单位面积苔藓植物生物量储量最高,平均910.10 g/m²,其次为腐木生(221.90 g/m²),树附生的最低(53.59 g/m²)。林窗地面生苔藓单位面积生物量最高,均值为360.47 g/m²,其次为林下(305.51 g/m²),最小为林缘(244.11 g/m²);林窗、林缘和林下间的地面生苔藓单位面积生物量差异显著(P＜0.05)。沿海拔梯度,林窗苔藓单位面积生物量表现出先增加后减少的趋势。因此,在西藏色季拉山暗针叶林内,林窗对地面生苔藓单位面积生物量有显著影响。     

广西北部湾海岸带、海岛的外来入侵植物

为保护广西北部湾海岸带、海岛的植物多样性和生态系统多样性,通过样方、样带法野外实地调查并结合文献资料,对其外来入侵植物的物种组成、原产地、生活型、入侵途径和危害状况等进行了分析。结果表明,广西北部湾海岸带、海岛共有入侵植物64种,隶属28科55属,其中菊科(Asteraceae)最多(15种)。草本植物最多,有48种(75.00%)。原产地来自美洲的植物最多,有49种。入侵风险等级可划分为5个等级,其中Ⅰ级严重危害的有8种(12.50%)。与广西、广东、海南及华南地区的外来入侵植物在物种组成、生活型和原产地等方面呈现出较强的相似性;北部湾与广西中越边境内陆地区来自美洲的入侵植物都超过60%。因此推测广西外来入侵植物有两条可能的入侵线路：一是从海南登录,二是从中越边境跨入。广西北部湾海岸带、海岛的外来入侵植物总数(相对整个广西)虽较少,但其8种Ⅰ级严重危害植物的防治,仍需引起重视。     

Phytochemical Characterization of Cannabis sativa L. Roots from Northeastern Brazil

This article describes the phytochemical study of Cannabis sativa roots from northeastern Brazil. The dried plant material was pulverized and subjected to exhaustive maceration with ethanol at room temperature, obtaining the crude ethanolic extract (Cs-EEBR). The volatile compounds were analyzed by gas chromatography coupled with mass spectrometry (GC/MS), which allowed to identify 22 compounds by comparing the linear retention index (LRI), the similarity index (SI) and the fragmentation pattern of the constituents with the literature. By this technique the major compounds identified were: friedelan-3-one and β-sitosterol. In addition, two fractions were obtained from Cs-EEBR by classical column chromatography and preparative thin layer chromatography. These fractions were analyzed by NMR and IR and together with the mass spectrometry data allowed to identify the compounds: epifriedelanol, friedelan-3-one, β-sitosterol and stigmasterol. The study contributed to the phytochemical knowledge of Cannabis sativa, specifically the roots, as there are few reports on the chemical constituents of this part of the plant.     

A chromosome-level genome assembly enables the identification of the follicule stimulating hormone receptor as the master sex-determining gene in the flatfish Solea senegalensis

Sex determination (SD) shows huge variation among fish and a high evolutionary rate, as illustrated by the Pleuronectiformes (flatfishes). This order is characterized by its adaptation to demersal life, compact genomes and diversity of SD mechanisms. Here, we assembled the Solea senegalensis genome, a flatfish of great commercial value, into 82 contigs (614 Mb) combining long- and short-read sequencing, which were next scaffolded using a highly dense genetic map (28,838 markers, 21 linkage groups), representing 98.9% of the assembly. Further, we established the correspondence between the assembly and the 21 chromosomes by using BAC-FISH. Whole genome resequencing of six males and six females enabled the identification of 41 single nucleotide polymorphism variants in the follicle stimulating hormone receptor (fshr) consistent with an XX/XY SD system. The observed sex association was validated in a broader independent sample, providing a novel molecular sexing tool. The fshr gene displayed differential expression between male and female gonads from 86 days post-fertilization, when the gonad is still an undifferentiated primordium, concomitant with the activation of amh and cyp19a1a, testis and ovary marker genes, respectively, in males and females. The Y-linked fshr allele, which included 24 nonsynonymous variants and showed a highly divergent 3D protein structure, was overexpressed in males compared to the X-linked allele at all stages of gonadal differentiation. We hypothesize a mechanism hampering the action of the follicle stimulating hormone driving the undifferentiated gonad toward testis.     

大肠杆菌rpoH基因的克隆,表达及其产物的纯化

本文用ＰＣＲ方法获得大肠杆菌热休克蛋白转录因子σ３２的编码基因ｒｐｏＨ,并克隆在含有ｔａｃ启动子的表达载体ｐＵＨＥ中,经ＩＰＴＧ诱导,在大肠杆菌中表达了Ｃ端融合有６个寡聚组氨酸的σ３２。表达产物经金属螯合层析一步纯化,达到ＳＤＳ－ＰＡＧＥ银染一条带纯度,氨基酸组成分析及Ｎ端序列分析结果与文献报道一致。３５Ｓ细胞内参入实验表明：即使在较低的温度下,表达产物σ３２（Ｈｉｓ）６也能导致热休克蛋白如ＧｒｏＥｌ、ＤｎａＫ、Ｈｔｐ的大量合成．     

新疆血革螨属一新种（蜱螨亚纲：血革螨科）

本文报道血革螨属一新种,命名为狭背血革螨,新种Ｈａｅｍｏｇａｍａｓｕｓａｎｇｕｓｔｕｓｓｐ．ｎｏｖ．采自新疆乌苏,寄主为野兔。