布鲁氏菌M5-90疫苗株virB2基因缺失株的构建及鉴定

【目的】构建布鲁氏菌M5-90疫苗株virB2基因缺失株。【方法】利用常规分子生物学技术构建自杀载体pGEM-7zf-ΔvirB2-sacB,通过同源重组的方法,将电转化后的布鲁氏菌分别经100 mg/L氨苄抗性筛选和5%蔗糖敏感性筛选,获得基因缺失株。对获得的基因缺失株进行PCR鉴定和稳定性检测。【结果】成功构建M5-90ΔvirB2基因缺失株,并且该缺失株在10代以内未发生回复突变。【结论】为研发新型布鲁氏菌弱毒基因缺失活苗奠定基础。     

'Representing' British Muslims: the strategic dimension to identity construction

This article explores how Muslim identity is constructed in different ways by two groups of political activists in Britain. At the heart of our investigation is an interest in how these different definitions of Muslim identity are organized to promote different forms of political action. We pay particular attention to how these groups employ the same Islamic concept, da'wah (the injunction to invite people to Islam), and show that it only gains meaning when invoked and deployed in the context of practical argumentation. That is, the meaning of such concepts is not a fixed given but highly contingent upon the contrasting strategic concerns of those claiming to represent the community. While these points have a general applicability, they are particularly important in the analysis of Muslim political activity. For too long, the ahistorical essentialist assumptions of 'Orientalism' have obscured the fundamentally contested and strategically constructed nature of Muslim identity.     

鼎湖山季风常绿阔叶林不同生境蕨类植物的叶片功能性状研究

为了解不同生境中蕨类植物的叶片功能,对鼎湖山季风常绿阔叶林中的6种优势蕨类植物的比叶面积、光合速率、元素含量、构建成本和水分利用效率等叶片性状进行了测定,研究了它们对环境的适应性。研究结果表明,两种附生蕨类(江南星蕨和崖姜蕨)的叶片具有较低的比叶面积(SLA)和光合速率(Aarea),较高的构建成本(CCarea)和碳同位素比率(?13C)。而两种林下土生蕨类(扇叶铁线蕨、桫椤)具有较高的Aarea和较低的?13C。生长在岩石基质上的中华里白具有最高的养分利用效率和较高的?13C。生长在溪流边的薄叶卷柏具有最高的SLA,最低的CCarea和?13C。这些揭示了蕨类植物叶片性状与生境之间具有紧密的相关性。由于附生蕨类的叶片具有高的能量投资和低的光合速率,对环境变化的适应能力弱,附生蕨类可能面临较大的生存压力。     

双表达骨形态发生蛋白2、9重组腺病毒载体的构建和表达

目的构建双表达骨形态发生蛋白（Bone morphogenic protein,BMP）2、9腺病毒重组体并进行鉴定。方法自单一表达的BMP2或BMP9 AdEasy质粒上扩增BMP2和BMP9片段,先后亚克隆至穿梭质粒pASG2,获得双表达穿梭质粒pASG2-BMP2、9。酶切及PCR鉴定确认、测序正确后同源重组获得双表达BMP2、BMP9腺病毒质粒,转染至HEK-293细胞中包装和扩增得到高滴度双表达BMP2、BMP9腺病毒,体外感染C3H10细胞,RT-PCR鉴定并观察其早期诱导成骨情况。结果成功构建双表达BMP2、BMP9的腺病毒,滴度约为1010IU/mL,RT-PCR证实双表达腺病毒在C3H10细胞中表达,其感染的C3H10细胞早期碱性磷酸酶含量较单一表达的BMP2或BMP9腺病毒组增加。结论成功构建双表达BMP2、9的重组腺病毒载体,为进一步研究BMP2和BMP9的协同成骨作用和制备高效的组织工程人工骨提供了有利的工具。     

asr基因酵母双杂交系统的构建及筛选鉴定

为研究asr作用的分子机制,分析该基因表达产物在体内相互作用的靶蛋白,分别构建诱饵质粒载体和转基因拟南芥cDNA文库,通过转入酵母细胞中,初步的鉴定分析证实成功构建了诱饵质拉载体和拟南芥cDNA文库.构建的酵母双杂交系统为发现asr基因表达产物在体内相互作用的靶蛋白奠定了基础.     

腺病毒介导的人抑瘤素M基因对A375人黑色素瘤细胞的抑制作用

目的:构建抑瘤素M(OSM)重组腺病毒载体,研究其对人黑色素瘤细胞A375的抑制作用。方法:以PEGZ-OSM重组质粒为模板,通过PCR技术扩增出OSM片段,采用腺病毒载体的基因重组和体外包装技术获得表达与人OSM氨基酸序列相同的重组腺病毒子Ad-OSM,感染A375细胞,用荧光显微镜、RT-PCR、Western blot法检测OSM在A375细胞中的转录和表达;荧光显微镜观察A375细胞的形态学改变;MTT法和流式细胞术(FCM)检测Ad-OSM对A375细胞的生长抑制和细胞周期的抑制效应;半定量RT-PCR法检测OSM基因表达对A375细胞中的Bax、Bcl-2基因表达的影响。结果:基因测序和PCR分析结果显示,成功构建了Ad-OSM腺病毒表达载体;RT-PCR和Western blot法检测到OSM基因在A375细胞中的转录和表达;OSM基因的表达对A375细胞增殖有明显抑制作用,并可诱导细胞凋亡,OSM基因可通过上调细胞中Bax和下调Bcl-2基因表达诱导细胞凋亡。结论:成功构建了Ad-OSM腺病毒表达载体,感染OSM基因可明显抑制A375人黑色素瘤细胞的生长,诱导其凋亡,该现象可能是通过改变Bax、Bcl-2基因表达水平来发挥抗肿瘤作用。     

探讨医院图书馆数字化信息建设与服务

数字化信息时代,信息资源建设是主流的业务。本文通过对医院图书馆数字资源建设与知识服务的内容,提出医院图书馆信息资源数字化建设的基本对策和发展方向。     

蝴蝶兰LFY基因克隆及高效植物表达载体的构建

根据NCBI中蝴蝶兰LFY花序分生组织基因序列设计2对引物,用RT-PCR法从蝴蝶兰花芽中扩增出LFY基因,对扩增产物进行克隆和测序.结果表明,获得的蝴蝶兰LFY基因约为1500 bp,与报道序列同源性达98.71％.将LFY基因插入pRI101-ON载体中,经PCR、双酶切及测序鉴定,证实重组表达质粒中含有目的片段,表明成功构建了高效植物表达载体pRI1O1-LFY.     

An attempt to dendrochronologically date house features at the King site (9FL5), a 16th century Late Mississippian town in northwestern Georgia,USA

The King site is a Late Mississippian (ca. 1400–1540 CE) aboriginal town located in northwestern Georgia along the Coosa River associated with the Coosa Chiefdom. The site was settled ca. 1530 but was occupied for perhaps only 50 years or so based on the lack of horizontal stratigraphy. The site was visited by members of either or both the Hernando de Soto expedition in 1540 and/or the Tristan de Luna expedition in 1560. In 1974, archaeologists discovered and removed 36 sections of subterranean charred pine posts from six house features. Our objectives were to determine if the tree rings on these posts could be dendrochronologically dated to verify the dates of site occupation and confirm the construction sequence of several houses determined originally via stratigraphic and archaeological evidence. We were able to graphically and statistically crossmatch 13 measurement series representing 10 posts from 5 of the 6 structures, yielding a 157-year floating chronology (average interseries correlation = 0.60). We were unable to absolutely crossdate this floating chronology with the only regional reference chronology long enough (back to 1378 CE) to reach the 16th century, an eastern red cedar chronology from eastern Tennessee. Archaeological evidence indicated Houses 8 and 23.4 were built later in the King site occupancy, confirmed by the tree-ring dates as both houses have the youngest tree rings of the five structures. House 14 had the oldest outermost tree rings but archaeological evidence suggests this house also was likely constructed late in the King site occupancy. We propose some posts were salvaged and reused from abandoned houses as the King site became rapidly depopulated in the last 10–20 years of site occupancy, thus explaining the age of posts used in House 14. We urge archaeologists working in the Southeastern U.S. to consider developing a more formal process for exhuming and preserving charcoalized wood remains from archaeological sites so that these samples can be evaluated using dendrochronological techniques.     

猪线粒体控制区微卫星(mtMs)变异及其分布特征

猪线粒体DNA长度因D-loop中mtMs序列重复数变异而不同。为阐明猪mtMs变异及其在不同群体中的分布特征,本研究对4个藏猪群体、八眉猪、烟台黑猪及3个引入猪群体共164个个体mtDNA D-loop全序进行测定,并与GenBank中发表的相关序列进行比对,分析了其中的重复单元核苷酸变异、重复次数及其在各群体间的分布规律。结果表明,因重复单元“5’-TGCGTACACG-3’”第2~4和10位上碱基变异,猪mtMs形成了以其为核心序列的多种重复单元(R^A^R^G)组成的复合结构,重复数介于3~47之间。单一重复(R^A)组成的mtMs结构在各群体中表现出分布优势,而大多数复合结构(R^AR^B)分布在国外选育群体中。藏猪的mtMs复合结构多达8个,R^AR^CR^E为其特有,另有5个与八眉猪共享。中国野猪和韩国地方猪种也有其特有mtMs结构。丰富的mtMs变异和特有结构与长期适应进化有关,本研究为地方猪种遗传资源及适应性研究提供了标记工具和理论基础。