9 resultados para aînés

em Chinese Academy of Sciences Institutional Repositories Grid Portal


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Sin and Pr doped CeO2 and Ce6MoO15 based materials were synthesized by sol-gel method. The structure of the powders were characterized by X-ray diffraction (XRD), Raman spectra, field emission scanning electron microscopy(FE-SEM) and the electrical conductivity of the samples was investigated by AC impedance spectroscopy. By comparing the structure and electrical properties of different systems, it could be concluded that the electrical property of Ce6MoO15 based system is better than that of CeO2 system. The added Mo element resulted in the increase of gain size and improved the grain boundary conductivity notably below 600 degrees C, while the Pr dopant induced the smaller grain size and improved the grain boundary conductivity of the materials.

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基因型变异和表型变异的相互关系问题是尚未解决的生物学基本问题之一,而解决这个问题的一个有效方法是研究基因组中功能相关基因的分子进化和表型效应的相互关系。在本研究中我们利用圆叶牵牛花青素代谢途径(该途径合成了与花色形成有关的色素)来研究表型变异和基因型变异的进化关系,希望探讨两个问题:1、代谢途径上基因多态性在不同位点的表现程度如何? 2、在中国种群中传粉者对花色有无选择? 在对第一个问题的研究中调查了中国圆叶牵牛花青素代谢途径部分结构基因和调控基因的多态性。所研究的结构基因为途径的第一个关键酶基因CHS-D基因和较下游的ANS基因、UF3GT基因,以及调控基因W位点。其次是在温室中进行人工自交以获得所研究基因的纯合体。在对第二个问题的研究中,首先调查了野外圆叶牵牛不同花色的传粉者和传粉几率,其次是分析有色花个体与白花个体中的遗传变异程度。目前的研究结果表明CHS-D位点有3个等位基因,其中在中国的西南部地区发现了1个新的、在已报道的7个等位基因以外的等位基因;较下游的结构酶基因ANS位点存在有5个等位基因(在中国新发现3个等位基因);UF3GT位点目前只发现有2个已报道的等位基因。在中国的圆叶牵牛中,新发现调控基因W位点上一个有功能的等位基因Ipmyb2,它与已报道的等位基因Ipmyb1的差异主要存在于内含子部分,而与发生缺失(1个6bp的缺失和1个19bp的缺失)后无功能的等位基因ipmyb2(该基因造成白花)存在外显子部分的8个碱基的差异。野外传粉观察结果表明,白花个体与有色花个体受到传粉者介导的选择,传粉者偏爱有色花而歧视白花,而且与在美洲不同,在中国圆叶牵牛的主要授粉者为蛾类而不是蜂类,从野外观察统计的结果看蛾类的传粉效率要高于蜂类。以上结果表明:1)花青素代谢途径上酶基因的位点均有一定程度的多态,其中尤以ANS为最高;调控基因ipmyb2的核苷酸缺失造成该基因停止功能,使其纯合子的个体开白色花。2)授粉者所介导的花色表型选择确实在中国种群中发生,有可能进一步增加花色代谢途径上基因位点的遗传变异。

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水母雪莲(Saussurea medusa Maxim)为名贵珍稀中药材,其主要药用成分为类黄酮,尤其是3-脱氧类黄酮。目前关于雪莲的研究主要集中在采用细胞培养生产类黄酮等方面,但对于雪莲类黄酮生物合成的分子机制了解甚少,极大限制了这一珍贵资源的利用。本研究采用水母雪莲红色系愈伤组织及悬浮细胞为材料,构建cDNA文库,从中克隆水母雪莲类黄酮次生代谢中的相关基因并对这些基因进行了深入的生物信息学分析、转基因研究初步确定其功能,以期了解雪莲类黄酮次生代谢的分子机制,为提高类黄酮的合成奠定基础。主要结果如下: 1. 成功地构建了水母雪莲红色系愈伤组织与悬浮细胞cDNA文库,原始文库滴度达到4×106pfu/ml,扩增文库滴度接近1011 pfu/ml,重组率达98%。PCR检测插入片段,均在0.5kb到3kb之间,1kb以上占62%。从文库中检测到了chs、dfr及Myb转录因子SmP,文库覆盖度达到要求且为PCR筛选文库提供了可能。 2. 采用部分简并引物,通过RT-PCR克隆了水母雪莲查尔酮异构酶基因Smchi特异探针,并根据这一探针序列设计特异引物,采用TD-PCR法筛选cDNA文库,获得Smchi cDNA序列,全长831bp,编码一个232氨基酸残基的蛋白。根据cDNA序列克隆了Smchi DNA序列,结果表明Smchi基因无内含子。Smchi cDNA序列与翠菊chi基因高度同源,ORF区域同源性高达84%,但推测氨基酸序列则只有79.3%。Smchi mRNA具有复杂的二级结构。SmCHI具有典型的Chalcone结构域,其二级结构与苜蓿CHI蛋白十分相似,7个α-螺旋与8个延伸链由随机结构联系起来。但其活性中心的第三个关键氨基酸残基N115为M115所取代,这一取代可能导致该蛋白无生物活性,也可能使它具有一般CHI不同的功能。构建Smchi正义、反义真核表达载体,通过农杆菌介导导入烟草,获得转正义、反义Smchi基因的烟草。转基因烟草花色未改变,但叶片总黄酮发生了显著的变化,50%转正义基因烟草总黄酮含量显著提高,最高比对照提高6倍,70%转反义基因烟草总黄酮含量显著下降,最多达85.1%,初步证明Smchi具有功能,并能有效调控烟草类黄酮次生代谢。因此,SmCHI可能是不同于已知CHI的一类新的CHI蛋白,它催化的反应可能与花色素合成无关,其反应机制也可能有所不同。 3. 伴随Smchi的克隆获得了一个黄烷酮3-羟化酶类似基因Smf3h的cDNA,全长1334bp,编码一个343aa的蛋白。根据这一cDNA序列克隆了Smf3h DNA序列,全长1630bp,结果表明该基因由4个外显子和3个内含子组成。Smf3h mRNA具有十分复杂的二级结构。 推测蛋白氨基酸同源性分析表明,SmF3H属于2OG-FeII_Oxy家族,与同一家族的的颠茄H6H的同源性为45%,与拟南芥F3H的同源性为40%,但对SmF3H、典型F3H及典型H6H推测蛋白二级结构、活性中心关键氨基酸残基的位置与相对距离、软件进行功能预测分析,发现SmF3H与F3H更相似。构建Smf3h的正义与反义真核表达载体,通过农杆菌介导导入烟草,但只获得一批转正义基因的烟草,反义基因导致烟草不能再生而未获得转反义基因烟草。转基因烟草花色未改变,叶片总黄酮也与对照相似,初步确认Smf3h与烟草类黄酮生物合成无关,而是一个既不属于f3h也不属于h6h的功能未确定的新基因。 4. 采用与克隆Smchi基因相似的方法,从cDNA文库中克隆了SmP基因cDNA,全长969bp,编码一个256 aa的蛋白质。根据cDNA序列克隆了SmP基因的DNA序列,结果表明,SmP基因无内含子。SmP基因cDNA 一级结构及mRNA二级结构预测分析表明,该基因A+T含量很高(63%),所形成二级结构以A-T配对为主,其稳定性可能较差。SmP推测蛋白序列具有R2R3-Myb转录因子的典型特征,在N-端具有两个Myb DNA-binding Domain,其二级结构与鸡Myb转录因子1A5J十分相似,与其他基因如水稻OsMYB、番茄ThMYB的同源区域主要集中在这一结构域,分别为71.3%和70.8%;C-端富含丝氨酸,与烟草NtMYB、葡萄VlMYB等类黄酮调控因子相似,都呈寡聚体分布,并具有相同的保守磷酸化位点S170与S206。构建SmP基因真核表达载体,通过农杆菌介导导入烟草,获得大量转基因烟草。转基因烟草花色未发生改变,但51%的转基因烟草叶片总黄酮含量都显著提高(0.5-6倍),表明SmP具有促进烟草类黄酮生物合成的功能,但所调控的支路与花色素合成无关。初步试验结果表明,转SmP基因烟草对蚜虫具有很高的抗性,可有效地抑制蚜虫在烟草上的生长,抑制率最高可达92%-100%。这一抗性与烟草中类黄酮的积累可能具有直接的联系,但还需要进一步的试验证明。 5. 与美国俄亥俄州立大学Erich Grotewold 博士实验室合作,完成了微型EST库50个克隆的测序并进行了分析,从中获得了水母雪莲花色素合酶基因SmANS及醛脱氢酶基因SmALDH的特异探针。根据SmANS特异探针设计引物,采用PCR从这50个克隆中筛选获得了SmANS的cDNA序列,全长1229bp,编码一个356aa的蛋白质。SmANS在cDNA水平上与同属的翠菊ANS基因高度同源,但同源区域集中在ORF区域,达到80%,mRNA 预测二级结构十分复杂;推测氨基酸序列与翠菊ANS同源性达到82.9%。SmANS属于2OG-FeII_Oxy家族,在2OG-FeII_Oxy结构域高度保守,与翠菊、甜橙ANS保守结构域同源性达到94%。预测蛋白二级结构以α-螺旋-β-折叠为主,由7个主螺旋和11个主β-折叠及随机结构连接而成,并具有2OG-FeII_Oxy家族活性中心的三个保守的组氨酸残基(His84、His235、His291)和一个天冬氨酸残基(Asp237)。 6. 根据微型EST库中获得的SmALDH特异探针设计引物,采用PCR从这50个克隆中筛选获得了SmALDH基因cDNA 序列,全长1664bp,编码一个491aa的蛋白质。SmALDH基因cDNA具有独特的碱基组成,3/-UTR富含A+T,占该区域碱基总量的80%,5/-UTR的A+T和G+C各占50%,比ORF区域(52%)还低,因此其mRNA二级结构中5/-UTR可以单独形成自身二级结构并且十分稳定,这可能影响基因的表达。这一现象在水稻、玉米等植物中也存在。SmALDH在cDNA水平上在ORF区域与拟南芥、藏红花、水稻等具有较高同源性,分别为64.03%、63.89%、63.72%,但在推测蛋白氨基酸序列水平上同源性反而较低,分别为54.9%、54.3%、54.0%。SmALDH缺少线粒体定位信号,为胞质醛脱氢酶,具有一个Aldedh 保守结构域,还具有与1OF7-H相似的以α-螺旋-β-折叠为主的二级结构,由10个主螺旋和15个主β-折叠及随机结构连接而成。由于ALDH在植物细胞乙醇发酵中具有解除醛类物质毒害的功能,因此SmALDH基因的克隆为改造细胞自身以适应发酵培养条件,解决水母雪莲细胞大规模培养中需氧问题提供了可能。

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p21Waf1/Cip1,是目前具有最广泛激酶抑制活性的细胞周期抑制蛋白。随着研究的发展,发现它在细胞增殖、分化、凋亡等方面具有越来越多的功能,并且这些功能很大程度上依赖于它能够与各种各样的靶蛋白相互作用。p21与靶分子的作用与它的结构特点密切相关, 因而从分子水平上研究p21蛋白的构象特点、以及其与靶分子的相互作用具有非常重要的意义。 在本文中,我们首先运用分子克隆技术在大肠杆菌中成功地构建了由温度控制的pBV220-p21表达体系。经30℃培养、42℃诱导表达以后,p21蛋白以包涵体形式得到高效表达,通过离子交换和分子筛两步纯化得到电泳纯的、具有生物活性的p21蛋白。利用生物化学、生物物理学等方法和手段我们首次较为系统地表征了p21蛋白的构象特点, 发现其具有典型的无规卷曲的二级结构;色氨酸很大程度上暴露于溶液中,处在正电荷的微环境中;首次发现p21蛋白能够与一些阴离子配合物(如suramin、Bis-ANS等)以近10-7数量级的解离常数直接相互作用,化学计量比为:1:1,进一步研究表明suramin在p21上的结合位点可能是p21的C-末端。在此基础上,我们通过丹磺酰氯标记过的钙调蛋白(CaM)的荧光光谱、p21141-164突变的色氨酸的内源荧光光谱及荧光猝灭方法、圆二色谱、非变性蛋白电泳等方法和手段表征了p21及p21141–164与CaM的作用。首次发现p21与CaM结合的解离常数在10-7数量级,p21与CaM之间除了疏水作用,还有较强的静电作用,以及一定的H键作用。深入研究表明p21141-164在CaM上的结合位点在CaM的Ca2+诱导的疏水区域,p21141–164的150和151位突变的色氨酸在结合后离CaM的疏水区域较近,而159位的色氨酸离的较远。同时发现p21141–164与CaM结合后只诱导出约14%的α螺旋结构,即p21141–164基本上是以无结构的状态与CaM结合的。

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The cloud-point temperatures (T-c1's) of ti-ans-decahydronaphthalene (TD)/polystyrene (PS, M-w = 270 kg/mol) solutions were determined by fight scattering measurements over a range of temperatures (1-16 degreesC), pressures (100-900 bar), and compositions (4.2-21.6 vol% polymer). The system phase separates upon cooling and the T-c1 was found to increase with the rising pressure for the constant composition. In the absence of special effects this finding indicates positive excess volumes. The special attention was paid to the demixing temperatures as a function of the pressure for the different polymer solutions and the plots in the T-volume fraction plane and P-volume fraction plane. The cloud-point curves of polymer solutions under changing pressures were observed for different compositions, demonstrates that the TD/PS system exhibits UCST (phase separation upon cooling) behavior. With this data the phase diagrams under pressure were calculated applying the Sanchez-Lacombe (SL) lattice fluid theory. Furthermore, the cause of phase separation, i.e., the influence of Flory-Huggins (FH) interaction parameter under pressure was investigated.

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Synchocystis sp. PCC 6803 lacks a gene for the any known types of lycopene cyclase. Recently, we reported that sll0659 (unknown for its function) from Synechocystis sp. PCC6803 shows similarity in sequence to a lycopene cyclase gene-CruA from Chlorobium tepidum. To test, whether Sll0659 encoded protein serves as lycopene cyclase, in this study, we investigated the carotenoids of the wild types ans mutants, In the sll0659 deleted mutant, there is no blockage at the lycopene cyclization step. Our results demonstrate that sll0659 does not affect lycopene cyclization. However, the ultrastructure of mutants suggests the involvement or necessity of sll0659 in the cell division.

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Glaciers in west China are the sources of the major great rivers in Asia, and the solid water resources are crucial to China and South Asia. Black carbon (BC) results in very complex climate effects not only in the atmosphere, but accelerates the melting after its deposit on the surface of snow/ice. As the main distributed area of glaciers in China, the Tibetan Plateau (TP) and Xinjiang region are abutted by South Asia, Central Asia, and Russia, and east China, and the atmospheric environment would be influenced by the BC emitted from these regions. Whereas, the BC’s temporal and spatial distributions for concentration in the mid and top troposphere in west China, its transport, and its radiative forcing after deposited on the snow/ice surface are not well understood at the present. In the field, we collected samples from surface snow, snow pits, ice core, and aerosol in the glaciers, analyzed BC content mainly by the thermo-oxidized method in the laboratory, and discussed temporal and spatial distributions for BC concentrations in glaciers, the transport, and its impacts on the environment. Several conclusions were derived as follows: 1_Spatial distribution and the impact on albedos for BC concentrations in snow/ice: the BC concentrations in the surface snow for the investigated glaciers could be placed in areas, the Tianshan Mountains > the central TP > the Pamirs > the Qilian Mountians > the Himalayas. This distribution could be attributed to the elevation of the glaciers, the topography of the TP, and more regional emissions. Probably significant impacts on the albedos of the glacier surface could be caused by BC deposits, and the estimated reduced albedos on the glaciers are 9.8% (the Zhadang glacier), 8.7% (the Miao’ergou Riverhead No.3 glacier), and 6.8% (the Kuitun River Haxilegen No.48 glacier), and 6.2% (the Dongkemadi glacier), and 5.3% (the La’nong glacier), and 4.2% (the Muztagata glacier), etc. 2_The temporal variance of BC concentrations in ice of the East Rongbuk Glacier (ERG) and its climatic implications: major cations and anions (e.g., SO42- and Ca2+) concentrations in aerosols during summer monsoon seasons showed their close relationships with the sources of air masses, in which the variance of SO42- concentrations suggested the atmospheric environment over the ERG was significantly influenced by the aerosols from South Asia. BC record based on an ice core suggested its deposit was dominantly transported by monsoons in summers and by westerlies in other seasons, and the BC from South Asia in summers dominated the varying trend of its concentrations in the ice core and caused higher concentrations in summers than those in other seasons. In the past 50 yrs, BC concentrations showed fluctuations, whereas showed an increasing tread in the most recent decade, and exceeded 50 μg kg-1 in the summer of 2001; correspondingly, the radiative forcing caused by BC showed an increasing trend since 1990s, and exceeded 4.5 W m-2 in the summer of 2001. 3_Cabonaceous aerosols in the Nam Co region: organic carbon (OC) concentration accounted for ~95% and BC for ~5% in the total carbonaceous aerosol concentration, which was significantly influenced by summer precipitations. OC was dominantly derived from fossil fuel burning and BC from both fossil fuel and biomass burning. Trajectory analysis and aerosol optical depth suggested the atmospheric environment in the Nam Co region was most probably influenced by the emissions from South Asia. The potential source regions of air pollutants in the Nam Co regions in summers might be Bangladesh and east India, and in winters might be the Indo-gangetic basin. The scavenging ratio of atmospheric BC by rainfalls was less than those at other sites. West China is a less-developed region for industry, where BC concentrations in the atmosphere and snow/ice could be significantly influenced by the emissions from the abutted regions with rising industries (South Asia, Central Asia, and Russia). For example, snow/ice BC concentrations in the glaciers of the Parmirs, the Tianshan Mountains, and the Qilian Mountains in the northeast margin of the TP might be more influenced by the emissions from Centrial Asia (transported by the westerlies), those in the glaciers of the Himalayas might be more influenced by the emissions from South Asia (transported by the monsoons and the westerlies), and atmospheric carbonaceous aerosols might also be more influenced by the emissions from South Asia (transported by the monsoons and the westerlies). The BC concentrations in some glaciers might cause significant impacts on the albedos for the glaciers, and therefore enhanced the radiative forcings, for example, the ERG. The research on the relationships among atmospheric and snow/ice BC and its radiative forcing, variance of snow cover, mass balance of glaciers, and climate forcing would be needed in future.

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These are two parts included in this report. In the first part, the zonation of the complexes in its series, lithofacies, the depth of magma source and chambers is discussed in detailed for the first time based on the new data of petrol-chemistry, isotopes, tectono-magma activity of Mesozoic volcano-plutonic complexes in the southern Great Hinggan Mts. Then, the genetic model of the zonality, double overlapped layer system, is proposed. The main conclusions are presented as follows: The Mesozoic volcanic-plutonic complexes in the southern Great Hinggan were formed by four stages of magma activity on the base of the subduction system formed in late Paleozoic. The Mesozoic magmatic activity began in Meso-Jurassic Epoch, flourished in late Jurassic Epoch, and declined in early Cretaceous Epoch. The complexes consist dominantly of acidic rocks with substantial intermediate rocks and a few mefic ones include the series of calc alkaline, high potassium calc alkaline, shoshonite, and a few alkaline. Most of those rocks are characterized by high potassium. The volcano-plutonic complexes is characterized by zonality, and can be divided mainly into there zones. The west zone, located in northwestern side of gneiss zone in Great Xinggan mountains, are dominated of high potassium basalts and basaltic andesite. The middle zone lies on the southeast side of the Proterozoic gneiss zone, and its southeast margin is along Huangganliang, Wushijiazi, and Baitazi. It composed of dominatly calc-alkaline, high potassium calc-alkaline rocks, deep granite and extrusive rhyolite. The east zone, occurring along Kesheketong Qi-Balinyou Qi-Balinzuo Qi, is dominated of shoshonite. In generally, southeastward from the Proterozoic gneiss zone, the Mesozoic plutons show the zones-mica granitites zone, hornblende-mica granitite zone, mica-hornblende granitite zone; the volcanic rocks also display the zones of calc alkaline-high potassium calc alkaline and shoshonites. In the same space, the late Paleozoic plutons also display the same zonality, which zones are combined of binary granite, granodiorite, quartz diorite and diorite southeast wards from the gneiss. Meso-Jurassic Epoch granite plutons almost distribute in the middle zone on the whole. Whereas late Jurassic Epoch volcanic rocks distribute in the west and east zone. This distribution of the volcano-plutonic complexes reveals that the middle zone was uplifted more intensively then the other zones in Meso-Jurassic and late Jurassic Epoches. Whole rock Rb-Sr isochron ages of the high potassium calc-alkaline volcanic rocks in the west zone, the calc-alkaline and high potassium calc-alkaline granite the middle zone, shoshonite in the east zone are 136Ma, 175Ma and 154Ma, respectively. The alkaline rocks close to the shoshonite zone is 143Ma and 126Ma. The isochron ages are comparable well with the K-Ar ages of the rocks obtained previously by other researchers. The compositions of Sr ans Nd isotopes suggest that the source of Mesozoic volcanic-plutonic complexes in Great Hinggan Mts. is mostly Paleo-Asia oceanic volcanic-sedimentary rocks, which probably was mixed by antiquated gneiss. The tectonic setting for Mesozoic magmatism was subductive continental margin. But this it was not directly formed by present west Pacific subduction. It actully was the re-working of the Paleozoic subduction system( which was formed during the Paleo-Asia ocean shortening) controlled by west Pacific subduction. For this reason, Although Great Hinggan Mts. is far away from west Pacific subduction zone, its volcanic arc still occurred echoing to the volcanic activities of east China, it, but the variation trend of potassium content in volcano-plutonic complexes of Great Hinggan is just reverse to ones of west Pacific. The primitive magmas occurred in the southern Great Hinggan Mts. Include high-potassium calc-alkaline basalt, high potassium calc-alkaline rhyolite, high potassium rhyolite, non-Eu negative anomaly trachy-rhyolite et al. Therefore, all of primitive magmas are either mafic or acid, and most of intermediate rocks occurring in the area are the products of Mesozoic acid magma contaminated by the Paleozoic volcanic- sedimentary rocks. The depth of those primitive magma sources and chambers gradually increase from northwest to southeast. This suggests that Paleozoic subduction still controlled the Mesozoic magmatism. In summary, the lithosphere tectonic system of the southern Great Hinggan Mts. controlling Mesozoic magmatism is a double overlapped layer system developing from Paleozoic subduction system. For this reason, the depth of crust of the southern Great Hinggan Mts. is thicker than that of its two sides, and consequently it causes regional negative gravity abnormity. The second part of this report shows the prolongation of the research work carried on in my doctor's period. Author presents new data about Rb-Sr and Sm-Nd isotopic compositions and ages, geochamical features, genesis mineralogy and ore deposit geology of the volcanic rocks in Kunyang rift. On the base of the substantial work, author presents a prospect of copper bearing magnetite ore deposit. The most important conclusions are as follows: 1. It is proved that all of these carbonatites controlled by a ringing structure system in Wuding-Lufeng basin in the central Yunnan were formed in the Mesoproterozoic period. Two stages could be identified as follows: in the first stage, carbonatitic volcanic rocks, such as lavas(Sm-Nd, 1685Ma), basaltic porphyrite dykes(Sm-Nd, 1645Ma), pyroclastic rocks and volcaniclastic sedimentary rocks, formed in the outer ring; in the second stage, carbonatitic breccias and dykes(Rb-Sr, 1048 Ma) did in the middle ring. The metamorphic age of the carbonatitic lavas (Rb-Sr, 893 Ma) in the outer ring was determined. The magma of carbonatitic volcanic rocks derived mainly form enriched mantle whose basement is depleted mantle that had been metasomated by mantle fluid and contaminated by Archaean lower crust. Carbonatitic spheres were discovered in ore bearing layers in Lishi copper mining in Yimen recently, which formed in calcite carbonatitic magma extrusion. This discovery indicates that the formation of copper ore deposit genesis relates to carbonatitic volcanic activity. The iron and copper ore deposits occurring in carbonatitic volcanic- sedimentary rocks in Kunyang rift results from carbonatitic magmatism. Author calls this kind of ore deposits as subaqueous carbonatitic iron-copper deposit. The magnetic anomaly area in the north of Lishi copper mining in Yimen was a depression more lower than its circumference. Iron and copper ores occurrig on the margin of the magnetic anomaly are volcanic hydrothermal deposit. The magnetic body causing the magnetic anomaly must be magnetite ore. Because the anomaly area is wide, it can be sure that there is a large insidious ore deposit embedding there.