Highly efficient soluble expression and purification of recombinant human basic fibroblast growth factor (hbFGF) by fusion with a new collagen-like protein (Scl2) in Escherichia coli

Abstract

Human basic fibroblast growth factor (hbFGF) is involved in a wide range of biological activities that affect the growth, differentiation, and migration. Due to its wound healing effects and therapy, hbFGF has the potential as therapeutic agent. Therefore, large-scale production of biologically active recombinant hbFGF with low cost is highly desirable. However, the complex structure of hbFGF hinders its high-level expression as the soluble and functional form. In the present study, an efficient, cost-effective, and scalable method for producing recombinant hbFGF was developed. The modified collagen-like protein (Scl2-M) from Streptococcus pyogenes was used as the fusion tag for producing recombinant hbFGF for the first time. After optimization, the expression level of Scl2-M-hbFGF reached approximately 0.85?g/L in the shake flask and 7.7?g/L in a high cell-density fermenter using glycerol as a carbon source. Then, the recombinant Scl2-M-hbFGF was readily purified using one-step acid precipitation and the purified Scl2-M-hbFGF was digested with enterokinase. The digested mixture was further subject to ion-exchange chromatography, and the final high-purity (96%) hbFGF product was prepared by freeze-drying. The recovery rate of the whole purification process attained 55.0%. In addition, the biological activity of recombinant hbFGF was confirmed by using L929 and BALB/c3T3 fibroblasts. Overall, this method has the potential for large scale production of recombinant hbFGF.     

Alteration of protein homeostasis mediates the interaction of Pseudomonas aeruginosa with Staphylococcus aureus

Intracellular protein degradation is essential for the survival of all organisms, but its role in interspecies interaction is unknown. Here, we show that the ClpXP protease of Pseudomonas aeruginosa suppresses its antimicrobial activity against Staphylococcus aureus, a common pathogen co-isolated with P. aeruginosa from polymicrobial human infections. Using proteomic, biochemical, and molecular genetic approaches, we found that this effect is due to the inhibitory effects of ClpXP on the quorum sensing (QS) of P. aeruginosa, mainly by degrading proteins (e.g., PhnA, PhnB, PqsR, and RhlI) which are critical for the production of QS signal molecules PQS and C4-HSL. We provide evidence that co-culturing with S. aureus induces a decrease in the activity of ClpXP in P. aeruginosa, an effect which was also achieved by the treatment of P. aeruginosa with N-acetylglucosamine (GlcNAc), a widespread chemical present on the surface of diverse cell types from bacteria to humans. These findings extend the range of biological events governed by proteolytic machinery to microbial community structure, thus also suggesting that a chemical-induced alteration of protein homeostasis is a mechanism for interspecies interactions.     

神经递质的可视化荧光检测技术研究进展

神经递质是神经系统中至关重要的组成部分,神经递质释放的时间和空间变化是神经网络中信息处理的核心,可视化监测神经递质的生物传感器是探究各类生理和病理活动的重要工具。文中综述了近年来具有较高时间和空间分辨率的监测神经递质时空分布变化技术的研究进展,介绍了对谷氨酸、多巴胺、γ-氨基丁酸和乙酰胆碱这4类重要的神经递质的检测方法,并归纳总结了各类检测方法的基本原理和优缺点,为设计具有高时空分辨率的神经递质传感器提供一个较为系统的参考。     

2030年闽三角城市群土地利用变化对生态系统水源涵养服务的影响

生态系统水源涵养服务作为水资源得以持续的基础和保障,正持续遭受着人类活动的干扰和影响,土地利用变化作为主要的方式之一,对水源涵养的影响广泛而且深远。运用InVEST模型模拟了闽三角城市群2015年和2030年的水源涵养情景,发现到2030年闽三角城市群区域内水源涵养量总体会下降0.24×10⁸ m³;对比发现：土地利用变化对水源涵养的影响主要主要表现在变化面积、变化方向、作用强度以及面积补偿作用四个方面。首先面积变化方面,水源涵养量同用地类型的面积大小正相关,但二者的变化量并不正相关;其次变化方向方面,相比变化为城市生态系统和水域生态系统而言,变化为自然生态系统和农业生态系统的土地利用变化更有利于生态系统水源涵养;再次,土地利用变化对水源涵养产生作用的强度由强及弱以此为林地、其他土地、草地、农田、建设用地、水域及滩涂;最后,面积变化后的补偿作用方面,由于不同用地类型水源涵养能力和面积变化量的差异,由农田、林地、草地及其他土地面积下降导致的水源涵养量损失并不能通过建设用地、水域及滩涂用地类型面积的增加得以完全补偿。     

景感生态学在城市生态系统服务中的应用研究——以城市公园景观设计为例

城市生态系统服务和可持续发展是当前城市生态学研究的热点问题。景感生态学作为联系生态系统服务和可持续发展的桥梁,可作为研究城市生态系服务和可持续发展的一种有效途径。随着社会发展所伴随的人们经济生活的生活压力增大,城市居民的亚健康状态日益突出。城市公园作为城市生态系统的重要构成,其设计目的应考虑应对城市居民健康问题和促进人类精神文明建设方面的作用。以城市公园景观设计为例,从园路、建筑、植物、水体景观和小品等方面探讨景感生态学在城市公园景观设计中的应用价值。景感生态学作为探索城市公园景观设计的新思路,以实现生态效益和居民福祉的提升,丰富和提升城市公园的生态系统服务功能,从而有利于促进为人类当代和后代提供可持续的福祉,以期驱使人类行为和言行规律朝着对生态系统有益的方向演化,自觉维护和改善生态系统服务,从而可持续地保障城市生态系统服务。     

基于Landsat-8影像的沿海城市公园冷岛效应——以厦门为例

城市公园主要由水体和绿色植被等景观构成,其形成的公园冷岛（Park cool island,PCI）已成为减缓城市热岛效应的重要途径之一。采用景感生态学中的景感营造理念规划设计城市公园或优化公园空间结构有助于增强公园冷岛效应,进一步改善局地城市热环境。选择沿海城市——厦门市为研究对象,基于2013年8月的Landsat-8 OLI/TIRS遥感影像和Google Earth高分影像数据,首先提取了研究区15个公园的土地利用信息,采用改进的地表温度单通道算法,获取了城市公园地表温度数据,并采用缓冲区分析、景观格局指数和多元统计等方法定量分析公园内部平均地表温度、城市公园冷岛强度（即降温幅度）、公园冷岛影响距离（即对周边区域的降温范围）三者的主要影响因子,并利用景感生态学原理初步剖析了城市公园景感营造的原则。结果显示：（1）公园面积和公园建设用地面积是影响公园平均温度的关键因子（R²=0.915）,这两个因子与公园平均温度均呈显著非线性相关;公园面积存在阈值55 hm²左右;（2）公园冷岛PCI强度由公园绿地面积、公园建筑面积和面积-周长形状指数（P/A）三个因子决定（R²=0.911）;公园冷岛PCI强度与公园绿地面积呈显著非线性正相关,与公园建筑面积呈显著非线性负相关,与面积-周长形状指数（P/A）呈显著线性负相关;在确保公园绿地面积达到一定阈值时,应该尽量降低公园建筑面积,增加公园形状复杂度,有利于其PCI强度的增加,缓解城市热岛效应;（3）公园冷岛影响距离由公园面积和公园水体面积比例决定（R²=0.719）,公园冷岛影响距离与两者呈显著正相关;绿地公园中提高公园水体的面积比例,有助于增大公园冷岛的影响距离;（4）城市公园的规划与设计,需要运用景感生态学的原理,从缓解城市热岛效应的角度,充分考虑公园大小、形状、土地利用类型组成比例及空间配置等因素。     

多杀菌素对天敌昆虫白蛾周氏啮小蜂的毒力评估

[目的] 生物防治与化学防治相结合是害虫综合防治的重要策略,蛹寄生蜂白蛾周氏啮小蜂是一种重要的天敌昆虫,在美国白蛾的生物防控中起关键作用。本研究旨在评估农林生产中常用的广谱、低毒杀虫剂多杀菌素对白蛾周氏啮小蜂的安全性。[方法] 采用药膜法检测多杀菌素对小蜂的毒力,并检测亚致死浓度多杀菌素处理后,小蜂体内解毒酶活性的变化。[结果] 多杀菌素对小蜂无短期（1 h）触杀作用,但随施药时间延长,其对小蜂的毒杀作用增强,25 mg·L^-1多杀菌素施药2 h后,可诱导约18.67%小蜂个体死亡。施药4 h后,不同浓度多杀菌素对小蜂的毒杀作用差异最明显,低浓度（5 mg·L^-1）多杀菌素可诱导11.33%小蜂死亡,而中、高浓度多杀菌素（≥ 15 mg·L^-1）可诱导77.33%~88.00%个体死亡。施药6 h后,LC₅₀值达7.44 mg·L^-1,安全系数为0.31,属高风险性农药。此外,亚致死浓度多杀菌素可明显诱导小蜂羧酸酯酶活性提高,对乙酰胆碱酯酶与谷胱甘肽-S-转移酶活性具有剂量效应。[结论] 多杀菌素对白蛾周氏啮小蜂的毒力作用较强,在美国白蛾生防区需慎用。     

Land masses and oceanic currents drive population structure of Heritiera littoralis,a widespread mangrove in the Indo‐West Pacific

Phylogeographic forces driving evolution of sea‐dispersed plants are often influenced by regional and species characteristics, although not yet deciphered at a large spatial scale for many taxa like the mangrove species Heritiera littoralis. This study aimed to assess geographic distribution of genetic variation of this widespread mangrove in the Indo‐West Pacific region and identify the phylogeographic factors influencing its present‐day distribution. Analysis of five chloroplast DNA fragments’ sequences from 37 populations revealed low genetic diversity at the population level and strong genetic structure of H. littoralis in this region. The estimated divergence times between the major genetic lineages indicated that glacial level changes during the Pleistocene epoch induced strong genetic differentiation across the Indian and Pacific Oceans. In comparison to the strong genetic break imposed by the Sunda Shelf toward splitting the lineages of the Indian and Pacific Oceans, the genetic differentiation between Indo‐Malesia and Australasia was not so prominent. Long‐distance dispersal ability of H. littoralis propagules helped the species to attain transoceanic distribution not only across South East Asia and Australia, but also across the Indian Ocean to East Africa. However, oceanic circulation pattern in the South China Sea was found to act as a barrier creating further intraoceanic genetic differentiation. Overall, phylogeographic analysis in this study revealed that glacial vicariance had profound influence on population differentiation in H. littoralis and caused low genetic diversity except for the refugia populations near the equator which might have persisted through glacial maxima. With increasing loss of suitable habitats due to anthropogenic activities, these findings therefore emphasize the urgent need for conservation actions for all populations throughout the distribution range of H. littoralis.