拟南芥白化突变体心口的基因定位与分析

EMS30是拟南芥经甲基磺酸乙酯（EMS）诱变得到的白化突变体。该突变体的叶绿体结构存在严重缺陷,同时伴随叶绿素缺失。遗传分析显示EMS30突变体的突变表型受隐性单基因控制。采用图位克隆的方法对EMS30突变基因进行定位的结果显示,该基因位于拟南芥第一条染色体的分子标记F21M12和F14N23之间的96kb区间内,该区间包含25个基因。通过生物信息学分析发现,该区间内有3个基因定位在叶绿体或与叶绿体发育相关。这些结果有助于该基因的克隆,为阐释叶绿体发育提供线索。     

整合素亲和力调控的机制

整合素是一类重要的细胞表面粘附分子,是由α和β两个亚基组成的异源二聚体跨膜蛋白。整合素作为细胞内外的桥梁,一方面负责介导细胞与细胞、细胞与细胞外基质以及细胞与病原体的相互作用,另一方面可以双向传递跨膜信号,对于免疫反应、免疫细胞的组织定位、凝血、组织愈伤、癌细胞转移以及组织和器官的发育等都至关重要。整合素与配体的结合及其相关的信号转导是受到精确调控的,这个过程伴随着整合素的一系列构象变化。整合素的另外一个特性是其与配体的结合受到二价金属阳离子的调控。本文重点介绍了整合素功能与构象的关系以及金属离子调控整合素功能的分子机制。     

过量表达番茄类囊体膜抗坏血酸过氧化物酶基因（StAPX）提高了烟草种苗的抗氧化能力

为了探讨叶绿体类囊体膜抗坏血酸过氧化物酶（tAPX）与其抗氧化性的关系,从番茄叶片中分离了叶绿体类囊体膜抗坏血酸过氧化物酶基因（StA跚并转入到烟草中。以野生型（WT）、转正义StAPX烟草株系T3-3和T3-6为试材,测定了外源过氧化氢诱导的氧化胁迫条件下APX酶活性、过氧化氢酶（CAT）活性、过氧化氢（H2O2）含量、叶绿素荧光参数及叶绿素含量等。Northern杂交显示StAPX因的表达受外源H2O2氧化胁迫的诱导。氧化胁迫下转基因烟草的APX酶活性和清除H2O2的能力都显著高于野生型,并且转基因烟草比野生型具有更高的PSII最大光化学效率及叶绿素含量。结果表明,．刚尸舶勺过量表达有助于提高外源H2O2诱导的转基因烟草的抗氧化能力。     

Identification of blood-protein carriers of the CA 19-9 antigen and characterization of prevalence in pancreatic diseases

The current best serum marker for pancreatic cancer, CA 19-9, detects a carbohydrate antigen on multiple protein carriers. Better knowledge of the protein carriers of the CA 19-9 antigen in various disease states may lead to improved diagnostic tests. To identify proteins that carry the CA 19-9 antigen, we immunoprecipitated the CA 19-9 antigen from pooled sera and identified the associated proteins using MS. Among the high-confidence identifications, we confirmed the presence of the CA 19-9 antigen on Apolipoprotein B-100 by antibody arrays and Western blot and on kininogen, ARVCF, and Apolipoprotein E by antibody arrays. We characterized the frequency and levels of the CA 19-9 antigen on the four proteins across various patient groups (pancreatic cancer, pancreatitis, and healthy controls) using antibody arrays. Nearly, 10-25% of the subjects showed elevations of the antigen on each protein, but the elevations were not associated with disease state or total CA 19-9 levels. These results contribute to our knowledge of the carrier proteins of an important functional glycan and the rate at which the glycan is displayed. This work also demonstrates a strategy for using the complementary methods of MS and antibody microarrays to identify protein carriers of glycans and assess the diagnostic value of measuring glycans on individual proteins.     

FUNDC1 regulates mitochondrial dynamics at the ER–mitochondrial contact site under hypoxic conditions

In hypoxic cells, dysfunctional mitochondria are selectively removed by a specialized autophagic process called mitophagy. The ER–mitochondrial contact site (MAM) is essential for fission of mitochondria prior to engulfment, and the outer mitochondrial membrane protein FUNDC1 interacts with LC3 to recruit autophagosomes, but the mechanisms integrating these processes are poorly understood. Here, we describe a new pathway mediating mitochondrial fission and subsequent mitophagy under hypoxic conditions. FUNDC1 accumulates at the MAM by associating with the ER membrane protein calnexin. As mitophagy proceeds, FUNDC1/calnexin association attenuates and the exposed cytosolic loop of FUNDC1 interacts with DRP1 instead. DRP1 is thereby recruited to the MAM, and mitochondrial fission then occurs. Knockdown of FUNDC1, DRP1, or calnexin prevents fission and mitophagy under hypoxic conditions. Thus, FUNDC1 integrates mitochondrial fission and mitophagy at the interface of the MAM by working in concert with DRP1 and calnexin under hypoxic conditions in mammalian cells.     

宁夏黄河流域景观破碎化时空变化特征

基于RS和GIS技术,以土地利用数据为基础,采用有效粒度尺寸对宁夏黄河流域1985年和2010年的景观破碎化进行时空变化分析。结果显示:流域整体的有效粒度尺寸(meff)从1985年的6326.62 km2下降到2010年的2974.32 km2,破碎化程度呈显著加剧变化;从流域内部来看,景观破碎化程度最大的是黄左区间,破碎化程度最小的是苦水河和红柳沟;除引黄区间外,其余分区景观破碎化程度在25年间均有所加剧。特征尺度分析结果显示研究区景观破碎化空间变异分析的合适尺度为4500 m;景观破碎化指数的空间变异结果表明破碎化程度较大的区域面积明显多于破碎化程度弱的区域,景观破碎化空间异质性在25年间表现出明显的上升趋势。在海拔背景条件下,2400 m以下区域的景观破碎化程度较高,2400m以上区域破碎化程度较低,且随海拔的升高有降低的趋势;流域景观破碎化受人为干扰影响强烈,在人为干扰较强的1000—1500 m区段的景观破碎化程度最大;景观破碎化在时间上的变化受人为干扰影响产生的变化最为显著,由自然条件改变产生的影响有限。研究结果可为西北地区景观格局及景观破碎化的研究提供参考,并为区域景观格局优化和土地的有效管理提供依据。     

新鲜土壤样品储存温度和时间对不同形态氮素的影响

土壤氮素形态及含量具有重要的生态学研究意义,而土壤样品的储存对土壤氮素含量的准确测定有很大影响.为了选择合理的土壤样品储存方法,本研究以福建省建瓯市万木林保护区罗浮栲林土壤为研究对象,测定在不同温度(25、4和-20 ℃)、不同储存时间(0、7和30 d)下土壤铵态氮、硝态氮、总氮、可溶性有机氮、氨基酸氮含量和微生物生物量氮,以及冷冻后常温培养过程中的氮素含量.结果表明: 在7 d的储存时间内,除氨基酸氮以外,常温培养样品下其余的氮素含量均有所增加;与新鲜样品相比,冷藏、冷冻样品的所有氮素含量之间均无显著性差异,且氮素含量变化较常温培养下更加稳定.因低温储存样品有刺激氮矿化的效果,在30 d储存时间内,与新鲜样品相比,除可溶性有机氮外,冷藏、冷冻样品的所有氮素含量均显著升高;两种冷储存方法之间无显著差异.因此,新鲜样品带回实验室后应及时处理;如需要冷储藏,时间不要超过半个月.如果需要较长的储存时间,则需将样品放置于更低的温度(-40或-80 ℃).在对储存土壤样品进行培养试验之前,需要进行预培养处理.在预培养过程中,除硝态氮含量呈现先下降再迅速升高的趋势外,其余氮素均随着培养时间逐渐趋近于新鲜土壤样品含量,在培养一周左右恢复到与新鲜土壤样品氮含量最为接近的状态.结合已有研究,对野外取样和风干样品需要5～14 d的预培养,冷储存样品预培养时间不应少于一周.     

大数据时代杂合酶的设计及其新趋势

酶分子的高效性和稳定性是工业广泛应用的物质基础。利用分子生物学技术可以将不同酶分子通过串联、插入、翻译后融合等方式构建成符合工业需求的杂合酶,但应用中多结构域杂合酶在表达量与酶活等方面仍存在弊端,而基于特定蛋白质结构域的多功能设计成为新趋势。高通量测序技术的发展,使得生物学家正面临着爆炸式增长的大数据集。近年来"蛋白质功能区"概念的提出,拓宽了人们对蛋白质结构与功能组织层次的认知,功能区残基聚簇的协同演化可导致同一家族不同蛋白质功能的差异。基于海量大数据分析可以快速定位特定功能区以及协同进化的关键位点,再利用合成生物学技术就可实现多种功能残基在同一蛋白质中的精准嫁接,完成天然酶分子的再设计。这将是杂合酶技术发展的新阶段,也会成为生物大数据时代下蛋白质设计的新趋势。     

Discovery of selective EGFR modulator to inhibit L858R/T790M double mutants bearing a N-9-Diphenyl-9H-purin-2-amine scaffold

Based upon the modeling binding mode of marketed AZD9291 with T790M, a series of N-9-Diphenyl-9H-purin-2-amine derivatives were designed and synthesized with the purpose to overcome the drug resistance resulted from T790M/L858R double mutations. The most potent compound 23a showed excellent enzyme inhibitory activities and selectivity with nanomolar IC₅₀ values for both the single T790M and double T790M/L858R mutant EGFRs, and was more than 8-fold selective for wild type EGFR. Compound 23a displayed strong antiproliferative activity against the H1975 non-small cell lung cancer (NSCLC) cells bearing T790M/L858R. And it was less potent against A549 (WT EGFR and k-Ras mutation) and HT-29 (non-special gene type) cells, showing a high safety index.     

Novel benzodiazepines derivatives as analgesic modulating for Transient receptor potential vanilloid 1

A new series of derivatives of 3-(7-chloro-5-(2-fluorophenyl)-2-oxo-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)propanoic acid were designed and synthesized as analgesic modulating for Transient receptor potential vanilloid 1. They were investigated for TRPV1 antagonistic activity in vitro, analgesic activity and sedative activity in vivo and aqueous solubility. Preliminary studies identified 3-(7-chloro-5-(2-fluorophenyl)-2-oxo-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)-N,N-dimethylpropanamide(Compound 11), as a potent analgesic modulating for TRPV1 with potent activity and good aqueous solubility.