Cloning and Expression of the Leukotoxin Gene SH from Fusobacterium necrophorum

坏死梭杆菌白细胞毒素是坏死杆菌病的主要致病因子,白细胞毒素基因(lkT)是其编码基因。以分离到的国内牛源坏死梭杆菌fn(A)菌株f4基因组dnA为模板,应用PCr方法扩增白细胞毒素基因SH片段,克隆至PMd18-T载体上,以bAMHⅠ和HIndⅢ酶切的目的片段SH与相应酶切的PET32A载体连接构建PET32A-SH重组表达质粒,经转化E COlI bl21(dE3)后用IPTg进行蛋白诱导,SdS-PAgE检测重组蛋白表达情况。结果表明:扩增基因序列大小为1800bP,SdS-PAgE检测重组蛋白有效表达,表达得到大小为80.2kdA的目的蛋白,采用镍柱亲和层析方法纯化SH重组蛋白,获得了纯度达95%的重组蛋白;经WEST-Ern-blOT证实,该蛋白对抗坏死杆菌阳性血清具有反应活性。The leukotoxin of Fusobacterium necrophorum(FN) is considered to be one of the main virulence factors.The lkt gene encodes for FN.In this study,the SH fragment of lkt gene was amplified by PCR using the F4 genome as the template,which was isolated from the Chinese Fusobacterium necrophorum strain.The fragment was then cloned to the pMD18-T vector for sequencing.Thereafter,the SH fragment was subcloned into the multiple cloning sites of the pET32 to construct pET32a-SH recombinant plasmid,which was then trans-formed into E.coli BL21(DE3) with IPTG induction for expression.SDS-PAGE was used to analyze the recombinant protein.The results showed that the SH fragment of about 1800 bp was amplified and was about 80.2 kDa.The fusion protein was purified by Ni-NTA affinity chromatography under denature conditions,and their purity was above 95%.Western-blot analysis indicated the SH fragment had anti-genicity against Fusobacterium necrophorum.“十五”国家科技攻关子课题(2002BA518A04);吉林省科技发展计划项目(20070570);吉林市科技发展计划项目(200805

2018年安溪珠塔内窑调查报告

2018年5月至7月,厦门大学历史系考古专业、福建博物院文物考古研究所及安溪县博物馆等单位联合组成\"安溪古窑址调查队\",对安溪珠塔内窑进行了全面的调查,基本了解珠塔内窑古窑址分布与保存情况,采集了一批明末清初的青花瓷器、白瓷与窑具标本,并发现部分窑炉遗迹,为研究闽南地区青花瓷窑业发展提供了第一手资料

Successive modification of polydentate complexes gives access to planar carbon-and nitrogen-based ligands

以碳和氮为键合原子的多齿螯合物是配合物家族中非常重要的一类。具有更高齿数的平面构型NC螯合物实例相对较少，代表性的例子为四齿金属碳卟啉类化合物，这类螯合物以其独特的结构和丰富的物理化学性质引起广泛关注。然而平面五齿、六齿的NC螯合物由于几何构型“拥挤”导致合成难度高，该工作从含三元环内金属卡宾结构的CCCC型碳龙配合物出发，利用经典有机反应（炔烃对金属卡宾的插入反应），成功地实现了CCCCN/NCCCN型平面五齿螯合物的合成。这一研究为高配位型螯合物的合成提供了新思路并为平面五齿螯合物家族添加新成员。特别是，这些高配位型螯合物在可见光和近红外区域均有较好的吸收，表现出良好的光声成像、光热转换及声动力学性能。 该研究工作在张弘教授指导下完成，第一作者为iChEM博士后周小茜。该工作充分体现了多学科协同研究优势：相关化合物的合成、表征及理论计算工作由周小茜博士完成；声动力学性能研究由厦门大学公共卫生学院庞鑫博士及刘刚教授完成；光声成像研究由厦门大学公共卫生学院聂立铭教授完成。iChEM fellow卓庆德博士、博士生卓凯玥、陈志昕参与了部分实验工作。夏海平教授、香港科技大学林振阳教授和南京大学朱从青教授对研究工作给予了大力支持。【Abstract】Polydentate complexes containing combinations of nitrogen and carbon (N and C) ligating atoms are among the most fundamental and ubiquitous molecules in coordination chemistry, yet the formation of such complexes with planar high-coordinate N/C sites remains challenging. Herein, we demonstrate an efficient route to access related complexes with tetradentate CCCN and pentadentate CCCCN and NCCCN cores by successive modification of the coordinating atoms in complexes with a CCCC core. Combined experimental and computational studies reveal that the rich reactivity of metal-carbon bonds and the inherent aromaticity of the metallacyclic skeletons play key roles in these transformations. This strategy addresses the paucity of synthetic approaches to mixed N/C planar pentadentate chelating species and provides valuable insights into the synthesis of carbon-based high-coordinate complexes. Furthermore, the resulting complexes are the examples of organometallic species with combined photoacoustic, photothermal, and sonodynamic properties, which makes them promising for application in related areas.This research was supported by the National Natural Science Foundation of China (Nos. 21572185, 21561162001, and 81571744), the Research Grants Council of Hong Kong (N_HKUST603/15), the Excellent Youth Foundation of Fujian Scientific Committee (2018J06024), and the Fundamental Research Funds for the Central Universities (20720170065).该工作得到国家自然科学基金委、香港研究资助局、福建省自然科学基金、厦门大学校长基金的资助