EMRE is an essential component of the mitochondrial calcium uniporter complex
and binding partner, MICU2, has not been extensively characterized (5). Other proteins,including leucine-zipper EF-hand containing transmembrane protein 1 (LETM1),mitochondrial calcium uniporter regulator 1 (MCUR1), mitochondrial sodium calcium exchanger (NCLX), transient receptor potential 3 (TRCP3), and uncoupling protein 2 and 3(UPC2 and 3) are also crucial for mitochondrial calcium physiology, but their physical relation to the uniplex are unclear (10–14).We took a biochemical approach to fully characterize composition the uniporter complex.We stably expressed MCU tagged with the FLAG epitope at its carboxy terminus (MCU-FLAG) in human embryonic kidney (HEK)-293T cells. MCU-FLAG restored mitochondrial calcium uptake in cells in which MCU was depleted with RNAi, and even caused a gain-of-function phenotype compared to that of cells that expressed a control protein (Fig. S1A).MCU exists in a large protein complex when isolated by digitonin permeabilization and native gel electrophoresis of mitochondria from HeLa cells or mouse liver (3). Similarly, in HEK-293T cells that stably expressed MCU-FLAG, MCU migrated at ~480 kD (Fig. 1A).Immunoprecipitation of MCU-FLAG, but not that of a control protein, yielded a protein complex of comparable size (Fig. 1A). Hence, MCU-FLAG associates with the apparent uniporter holocomplex, which we call the uniplex (uni porter com plex ).To define the components of the uniplex, we used a quantitative mass spectrometry approach using stable isotope labelling by amino acids in cell culture (SILAC) (15). We grew MCU-FLAG expressing and wild-type HEK-293T cells in medium containing heavy or light amino acid isotopes, respectively, immunoprecipitated proteins with FLAG antibody-conjugated beads and mixed heavy and light samples before they were processed for mass spectrometry. Two independent quantitative mass spectrometry experiments, with
two different digestion protocols, reproducibly identified only five known or predicted
mitochondrial proteins (Fig. 1B, Table S1). MCU was enriched in MCU-FLAG samples, as well as its known regulators MICU1 and MICU2. The uniplex also contains MCUb, a
paralog of MCU (16). The only other mitochondrial protein revealed through this analysis was C22orf32, a previously uncharacterized protein we call essential mcu regulator
(EMRE). MICU1, MICU2, MCUb and EMRE were detected by protein immunoblotting after immunoprecipitation of MCU-FLAG from HEK-293T and HeLa cells, but not with succinate dehydrogenase iron-sulfur subunit SDHB-FLAG, a protein also localized to the inner membrane (Fig. 1C, S1B). Moreover, MCU overexpression resulted in increased abundance of MICU1, MICU2 and EMRE (Fig 1C), probably because it stabilized its binding partners.
EMRE is a ~10 kD protein with a predicted mitochondrial targeting sequence, a predicted transmembrane domain (17, 18), and a highly conserved C-terminus rich in aspartate
residues (Fig. 2A). MCU and MICU1 are found in all major eukaryotic taxa, with lineage specific losses in several clades. We did not find EMRE homologs in plants or protozoa.Analysis of recently sequenced opisthokonts and basal fungi revealed that EMRE homologs are not found in any fungi, indicting that it most likely arose in the metazoan lineage (Fig.2B). EMRE RNA was broadly expressed in all mouse tissues (Fig. S2). Similar to MCU and MICU1, EMRE was identified by proteomic analysis of mitochondria (19) with a pattern suggesting it is ubiquitously expressed in the mitochondria of all mammalian tissues. EMRE
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appears to be a bona fide transmembrane protein as it was resistant to carbonate extraction at high pH, as is MCU (Fig. 2C).We tested the impact of loss of EMRE on uniplex function by RNAi mediated silencing of EMRE. Depletion of EMRE with each of two, sequence-independent hairpins led to loss of mitochondrial calcium uptake equivalent to MCU silencing in permeabilized HEK-293T and HeLa cells, as well as in intact HeLa cells after histamine stimulation (Fig. 3A, S3A, B). The appearance of the cells and their rates of proliferation were normal after EMRE silencing.The mitochondrial membrane potential was intact and could be depolarized in cells depleted of EMRE (Fig. S3C), indicating that loss of calcium uptake was not a trivial consequence of loss of the mitochondrial membrane potential. Overexpression of MCU in cells depleted of EMRE failed to restore mitochondrial calcium uptake (Fig. 3B), suggesting that MCU is not sufficient for in vivo uniporter current as previously proposed (4).To characterize complete loss-of-function phenotypes of uniplex components, we generated HEK-293T cells lacking MCU, MICU1 or EMRE with TALEN technology (Fig. 3C).Surprisingly, in cells lacking MCU, abundance of EMRE was decreased compared to that of wild type cells (Fig. 3C). However, in these cells, abundance of EMRE mRNA was similar to that in wild type cells (Fig. S3D), suggesting that loss of EMRE occurred post-transcriptionally. Thus, EMRE may be destabilized when its binding partner MCU is lost,analogous to the dependence of MICU2 protein expression on MICU1 (Fig. 3C and (5)).Cells lacking EMRE, similar to cells lacking MCU, exhibited severe defects in mitochondrial calcium uptake (Fig. 3D, S3E). In cells lacking MICU1, depletion of EMRE also leads to loss of mitochondrial calcium uptake (Fig. S3F). Digitonin permeabilization of isolated mitochondria from control cell or cells lacking EMRE followed by proteolysis and
western blotting showed that abundance of other uniplex proteins and their mitochondrial localization did not change after loss of EMRE (Fig. 3C, 3E, S3G). These experiments confirmed the topology of MCU (3, 7) and MICU1 (9) and showed that the topology of
MCUb is similar to the topology of MCU and MICU2 localizes to the intermembrane space like MICU1 (Fig. 3E, SG). To show that loss of EMRE specifically affects the uniporter conductance and not other mitochondrial parameters that will impact mitochondrial calcium uptake (e.g., proton motive force, pH, buffering capacity), we voltage-clamped mitoplasts isolated from cells lacking EMRE or these same cells after stable expression of EMRE, and measured mitochondrial calcium current (I MiCa ). Mitoplasts from cells lacking EMRE has significantly reduced I MiCa whereas typical I MiCa was seen after EMRE re-expression (Fig.3F) (6).
Consistent with EMRE being a core component of the uniplex, when EMRE was lost, the size of the complex on a native gel reduced to ~300 kDa, similar to that in cells lacking MICU1 (Fig. 4A). The observation that loss of either EMRE or MICU1 reduced the size of the native complex to a similar extent raised the hypothesis that EMRE may mediate
interaction between MCU and MICU1 and MICU2. To test this, we stably expressed MCU-FLAG or a control protein in HEK-293T or HeLa wild-type cells or cells lacking EMRE and performed FLAG immunoprecipitations. In wild-type cells, MICU1 and MICU2
immunoprecipitated with MCU-FLAG, but not with control SDHB-FLAG (Fig. 4B, S4).However, in cells lacking EMRE, the interaction between MCU-FLAG and MICU1 and
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MICU2 was completely lost (Fig. 4B, S4). Similarly, MCU was not associated with immunoprecipitated MICU1-FLAG in cells lacking EMRE (Fig. 4C). In the absence of EMRE, MCU still oligomerized (Fig. 4A, B) and interacted with MCUb. Moreover, in the absence of EMRE, the interaction between MICU1 and MICU2 was intact (Fig. 4C). These data indicate that loss of EMRE specifically interrupted the association of MCU with
MICU1 and MICU2. Furthermore, MCUb, MICU1 and MICU2 appear to be dispensable for MCU-EMRE interaction because in cells lacking MCUb or MICU1, EMRE was still associated with immunoprecipitated MCU-FLAG (Fig. 4D, E).
We propose a model where EMRE interacts with MICU1 and MICU2 in the IMS and with MCU oligomers in the inner membrane, thus linking the calcium sensing activity of MICU1and MICU2 to the channel activity of MCU (Fig. 4F). The fact that our immunoprecipitated complex migrates at the same apparent molecular size as the uniporter from purified
mitochondria (Fig. 1A), and recovers the founding members of the complex, suggests that we immunoprecipiated all of its components. It is notable that although other proteins have been reported to participate in mitochondrial calcium handling, such as LETM1, NCLX,UCP2 and 3, MCUR1 and TRCP3, they were not recovered in our proteomic assay,
suggesting that they play key roles outside of the uniplex. A key insight from the current work is that EMRE is essential for in vivo uniporter current (I MiCa ) and that MCU oligomers alone are not sufficient for in vivo uniporter activity, in contrast to what has been suggested using in vitro bilayer studies (4). Why MCU needs association with EMRE for in vivo
calcium conductance requires further investigation. Phylogenetic analysis indicate that the uniporter must have been a feature of the earliest mitochondria, since MCU and MICU1 are found within all major eukaryotic taxa, with lineage specific losses (20). EMRE is unique amongst human uniplex components in that it appears to have emerged more recently and represents a metazoan innovation. Its identification should help us understand how the activity and regulation of this ancient channel evolved.Supplementary Material
Refer to Web version on PubMed Central for supplementary material.
Acknowledgments
We thank Z. Grabarek and D.M. Shechner for helpful discussions. Y.S. received support from Helen Hay Whitney Foundation. D.C. received support from NIH F32HL107021. D.E.C. is an Investigator of the Howard Hughes Medical Institute. This work was supported by grants to V.K.M from NIH DK080261 and a gift from W. Dan and Pat Wright.
References and Notes
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2. Perocchi F, et al. MICU1 encodes a mitochondrial EF hand protein required for Ca(2+) uptake.Nature. Sep 16.2010 467:291. [PubMed: 20693986]
3. Baughman JM, et al. Integrative genomics identifies MCU as an essential component of the mitochondrial calcium uniporter. Nature. Aug 18.2011 476:341. [PubMed: 21685886]
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4. De Stefani D, Raffaello A, Teardo E, Szabo I, Rizzuto R. A forty-kilodalton protein of the inner membrane is the mitochondrial calcium uniporter. Nature. Aug 18.2011 476:336. [PubMed:21685888]
5. Plovanich M, et al. MICU2, a paralog of MICU1, resides within the mitochondrial uniporter complex to regulate calcium handling. PLoS One. 2013; 8:e55785. [PubMed: 23409044]
6. Chaudhuri D, Sancak Y, Mootha VK, Clapham DE. MCU encodes the pore conducting mitochondrial calcium currents. Elife. 2013; 2:e00704. [PubMed: 23755363]
7. Martell JD, et al. Engineered ascorbate peroxidase as a genetically encoded reporter for electron microscopy. Nat Biotechnol. Nov.2012 30:1143. [PubMed: 23086203]
8. Mallilankaraman K, et al. MICU1 is an essential gatekeeper for MCU-mediated mitochondrial Ca(2+) uptake that regulates cell survival. Cell. Oct 26.2012 151:630. [PubMed: 23101630]
9. Csordas G, et al. MICU1 Controls Both the Threshold and Cooperative Activation of the Mitochondrial Ca(2+) Uniporter. Cell Metab. Jun 4.2013 17:976. [PubMed: 23747253]10. Palty R, et al. Lithium-calcium exchange is mediated by a distinct potassium-independent sodium-calcium exchanger. J Biol Chem. Jun 11.2004 279:25234. [PubMed: 15060069]11. Jiang D, Zhao L, Clapham DE. Genome-wide RNAi screen identifies Letm1 as a mitochondrial Ca2+/H+ antiporter. Science. Oct 2.2009 326:144. [PubMed: 19797662]12. Mallilankaraman K, et al. MCUR1 is an essential component of mitochondrial Ca2+ uptake that regulates cellular metabolism. Nat Cell Biol. Dec.2012 14:1336. [PubMed: 23178883]13. Feng S, et al. Canonical transient receptor potential 3 channels regulate mitochondrial calcium uptake. Proc Natl Acad Sci U S A. Jul 2.2013 110:11011. [PubMed: 23776229]14. Trenker M, Malli R, Fertschai I, Levak-Frank S, Graier WF. Uncoupling proteins 2 and 3 are fundamental for mitochondrial Ca2+ uniport. Nat Cell Biol. Apr.2007 9:445. [PubMed: 17351641]15. Ong SE, et al. Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics. Mol Cell Proteomics. May.2002 1:376. [PubMed:12118079]16. Raffaello A, et al. The mitochondrial calcium uniporter is a multimer that can include a dominant-negative pore-forming subunit. Embo J. Jul 30.2013
17. Krogh A, Larsson B, von Heijne G, Sonnhammer EL. Predicting transmembrane protein topology
with a hidden Markov model: application to complete genomes. J Mol Biol. Jan 19.2001 305:567.
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Fig. 1. Affinity purification and proteomic analysis of the uniporter complex, uniplex (A) MCU-FLAG or control SDHB-FLAG was stably expressed in HEK-293T cells. Proteins from digitonin-permeabilized mitochondria from MCU-FLAG expressing cells and FLAG
immunoprecipitations from SDHB-FLAG and MCU-FLAG expressing cells were subjected to Blue Native-PAGE (BN-PAGE) and immunoblotted with MCU antibody. (B)
Identification of proteins that interact with MCU-FLAG. MCU-FLAG expressing cells were grown in the presence of heavy amino acids, control HEK-293T cells were grown in the presence of light amino acids. FLAG immunoprecipitates from both samples were mixed and analyzed by mass spectrometry. The ratios of heavy and light proteins annotated with mitochondrial localization from two replicates are shown. Proteins that show significant enrichment in heavy samples are shown in red. (C) Interaction of MCUb, MICU1, MICU2and EMRE with MCU. MCU-FLAG or control SDHB-FLAG was immunoprecipitated from HEK-293T cells. Immunoprecipitates and cell lysates were analyzed by immunoblotting for the indicated proteins.
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Fig. 2. Domain architecture, phylogeny and membrane association of EMRE
(A) Schematics showing predicted mitochondrial targeting sequence (MTS), transmembrane (TM) and conserved carboxy terminal acidic domain (CAD) of EMRE. The CAD of EMRE from six species was aligned using BLOSUM similarity matrix. (B) EMRE is metazoan-specific. Presence of homologs of MCU/MCUb, MICU1/MICU2 and EMRE across 20selected species spanning the NCBI taxonomy tree, indicating that EMRE is a metazoan innovation. (C) EMRE is a membrane protein. HEK-293T cell mitochondria were isolated and proteins were extracted with 0.1M Na 2CO 3 at pH 10 and pH 11.5. EMRE is observed in the insoluble pellet (P) similar to MCU. Cytochrome c (CYCS) is loosely associated with the inner membrane and is detected in the soluble fraction (S).
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Fig. 3. Requirement of EMRE for uniporter activity
(A) EMRE knock down impairs mitochondrial calcium uptake. Digitonin permeabilized HEK-293T cells were incubated with the calcium indicator Oregon Green Bapta 6F (OGB6F). After addition of 50 μM CaCl 2 (arrow), the depletion of extracellular calcium due to mitochondrial uptake was monitored by OGB6F fluorescence. Representative traces of mitochondrial calcium uptake from HEK-293T cells with GFP (control), MCU and EMRE knock down are shown. The bar graph shows the rate of calcium uptake relative to shGFP cells (mean ± s.d., n=4). (B) MCU overexpression cannot rescue mitochondrial calcium uptake in EMRE knock down cells. EMRE was knocked down in HEK-293T cells that
overexpress MCU-FLAG. The samples were treated as in (A) (n=4). Lysates were analyzed by immunoblotting for indicated proteins. (C) Loss of MCU decreases EMRE and MCUb abundance. MCU, MICU1 and EMRE knockout (KO) HEK-293T cells were generated
using TALEN technology, cell lysates from wild type (WT) and two independent knockout clones were prepared and analyzed by immunoblotting for control ATP5A and uniplex proteins (D) Mitochondrial calcium uptake is severely impaired in cells that lack EMRE.Mitochondrial calcium uptake was measured in WT and two independent EMRE KO cell lines as in (A). (E) Loss of EMRE does not change MCU, MCUb, MICU1 and MICU2submitochondrial localization. Mitochondria were isolated from cells lacking EMRE,
incubated with increasing concentrations of digitonin in the presence of proteinase K (PK),samples were analyzed by immunoblotting for outer membrane protein TOMM20, inner membrane protein TIMM23, matrix protein HSP60, as well as uniplex proteins (F)
Exemplar trace (red) of a mitoplast derived from EMRE knockout cells demonstrates absent
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calcium current (I MiCa), whereas typical I MiCa is seen after EMRE is knocked back in
(black). Voltage ramps were delivered from ?160mV to +80mV for 750ms, using a holding
potential of 0mV. (Right) Summary data. Error bars report SEM. NIH-PA Author Manuscript
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Fig. 4. Requirement of EMRE for the interaction of MCU with MICU1 and MICU2(A) Loss of EMRE or MICU1 reduces the size of the uniplex to ~300 kD. Mitochondria from indicated cell lines were isolated, digitonin permeabilized and immunoblotted for MCU after BN-PAGE. EMRE mediates MCU-MICU1 and MICU2 interaction. Stably
expressed SDHB-FLAG (control) or MCU-FLAG (B) or MICU1-FLAG (C) was
immunoprecipitated from WT or EMRE knockout cells, lysates and immunoprecipitates were analyzed by immunoblotting for indicated proteins. (D) Loss of MCUb does not alter MCU-EMRE, MICU1 and MICU2 interaction. Stably expressed SDHB-FLAG or MCU-FLAG was immunoprecipitated from cells after GFP (control) or MCUb knock down with two different hairpins. Samples were treated and analyzed as in (B). (E) Loss of MICU1does not alter MCU-EMRE interaction. Plasmid for EMRE expression was co-transfected with plasmids for control mitochondrial GFP-FLAG or MCU-FLAG expression, in WT and MICU1 knockout cells. Samples were treated and analyzed as in (B). (F) Model showing submitochondrial localization and organization of uniplex proteins. MCU is the pore-
forming component of the uniplex; MICU1 and MICU2 reside in the IMS where they sense outside calcium and regulate MCU through EMRE. EMRE is a single-pass transmembrane protein. It bridges MCU and MICU1 and MICU2 and is essential for the activity of the uniporter. MCU-MCUb interaction is not mediated by EMRE.
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翻译项目的字数是影响收费的重要因素之一，翻译字数主要对于笔译而言，例如：文件翻译、图书翻译、资料翻译、画册翻译等等，这些文件资料的字数决定了项目的翻译价格和翻译收费标准。 5.根据翻译项目语种 主流语种：英语、日语、韩语等和小语种：阿拉伯语、希腊语、印尼语等的翻译收费标准区别。我们知道：“物以稀为贵，”所以小语种的翻译报价会比主流语种收费要高的。 6.根据翻译项目难易程度 对于翻译公司来说，翻译费在很大程度上取决于翻译的难度程度，不同的行业术语不同，难度不同; 专业翻译公司将根据翻译人员的翻译水平、专业知识、翻译经验等方式来评价自己的翻译团队，高层次的翻译人员当然都是高收费; 如通用翻译、精细翻译、出版层次等不同类型的翻译报价不同，稿件的行业领域、材料难度、选择翻译类型等都是决定翻译公司收费标准的因素。
英文合同翻译价格 英文合同翻译需要多少钱
英文合同翻译价格英文合同翻译需要多少钱 在企业的经营过程中，有时候可能会涉及到翻译这个问题，但是一般的小企业并没有专门的人去做这件事情，大部分都是外包。那么对于企业来讲，翻译一份英文合同需要多少钱呢？作为浙江省最大的翻译公司，以琳翻译就在这里为大家解读一下。 一般来讲，翻译这项服务都是以字数来计价的，市场上的一般的价格是50-80元/千字，这是一个基本的价格。但是不同的公司的专业性质不一样的话，所给出的价格也是不一样的。对于公司的衡量标准来讲，影响价格的因素主要有：公司的资历、翻译人员的专业性、翻译文件的种类、难度等。所以，如果你需要去找翻译公司去服务，那么就需要考虑这些方面的东西。而对于合同这种文件，对于公司来讲是十分重要的，所以也需要去找专业的公司去进行翻译，如果是找一个资质不够的公司或者团队，那么就可能产生一些意想不到的问题，从而影响到公司的最终利益。 下面，我们来看看以琳翻译给出的翻译的价格。 从上面的价格可以看出，以琳翻译给出的价格是高于一般市场上的价格的，最低级别的翻译是160元/千字，然后分为A、B、C三级。C级译稿为普通中籍译员+中籍译员审核，满足客户对译文的普通要求。这是对于一般的合同而言的，但是如果是部分专业性质较强或者要求比较高的译文的话，那么可以选择更高级别的翻译，当然价格还是相对比较高的。 那么以琳翻译的资质是怎么样呢？我们再来看一下。 杭州以琳翻译有限公司是浙江省最大的实体翻译公司、中国翻译协会单位会员、美国翻译协会会员、全国翻译专业硕士研究生教育实习基地、西博会指定合作伙伴、以琳杭州翻译公司翻译团队成员均具有五年以上专业翻译、项目管理经验，绝大部分成员具有十年以上行业翻译经验。翻译服务涵盖英语、法语、韩语、日语、德语、俄语、西班牙语、葡萄牙语、
专业英文翻译中文收费标准
精诚英语翻译报价50-80元千字（市场价格100左右 精诚英语翻译工作室是由众多英语方面精英组成的翻译团队,一直致力于为广大中小企业和个人提供专业低价中英文翻译服务。价格是我们永远的优势！！！！最低价格支付宝担保交易，让你省钱又放心接受试译！！自信源于专业可以百度搜索精诚英语翻译找到我们 选择我们的理由：可以百度搜索精诚英语翻译找到我们 1.保证价格最低，团队网络化运作，无需经营成本，可以通过低价让利于客户。（有些客户看到这么低的价格还不敢相信，但是对于我们来说是完全可以接受的。） 2.保证准时、保密、准确 3.接受淘宝交易，让您没有任何担忧。 4.长期翻译经验，保证质量让您满意。 龚如心遗产案虽然告一段落，「遗产」二字仍然成为近日香港的焦点。新春期间，民政事务局局长曾德成表示，政府将展开全港非物质文化遗产首期普查，希望市民为遗产清单提出建议。 「遗产」是「资产」？ 近五、六年间，香港对保护本地小区和文化传统的意识高涨，现在政府带头要列一个「非物质文化遗产」清单，理应是很受欢迎之举。不过普查尚未展开，就引来学者争议，其中单是「非物质文化遗产」这个译名，就引起不少误会。 「非物质文化遗产」的原文是intangible cultural heritage（英文）或patrimoine culturel immateriel （法文），是联合国在1997年以尊重多元文化为大原则而提出的概念，并由联合国教科文组织制定「保护非物质文化遗产公约」，2006年生效。 「非物质文化遗产」是中国大陆的翻译，香港有学者不约而同就「非物质」和「遗产」二字提出质疑。香港城市大学中国文化中心主任郑培凯早在2005年就大声疾呼译名不妥。他认为原文heritage/patrimoine的意义是「传承」而非资产，不容易引发出财产的概念。而现在约定俗成译作「遗产」，容易令人觉得祖宗留下的东西，是可以变卖和投资的生财工具，与联合国提出的文化传承精神背道而驰。郑教授认识，正确的译名是「非物质文化承继」或「非实物文化传承」。另一位民俗学研究者陈云进一步指出，intangible「乃触摸不到的事，无形无相之事」，应用「精神价值」代之，「非物质」有消灭了精神之嫌，所以中国人应堂堂正正将之翻译为「无形文化传承」。 姗姗来迟的「遗产」 不论是「非物质」还是「无形」，「遗产」还是「承传」，即使公约成员国中国曲译甚至错译，香港特区政府还是只能照单全收。而且，随之而来的不止是字面的斟酌，而是「遗产」的搜寻和管理问题。 中国自2005年起，就开始非物质文化遗产（由于这个名称已约定俗成，故下文仍沿用之，并简称为「非遗」）普查，并陆续列出清单。香港也在翌年提出编制非遗清单，但却延至去年才聘专家普查，估计最快要2012年才完成。 非遗普查尚未展开，在国家文化部的再三邀请（或是说催促？）下，去年九月，香港终于申请将长洲太平清醮、大澳端午游涌、大坑舞火龙和香港潮人盂兰胜会列为第三批国家级非遗，预计今年六月有结果。 其实香港已错过了2006和2008年首批和第二批的申报机会，所以，至目前为止，在中国的文化版图上，香港是唯一没有任何有形和无形「遗产」的主要城市／特区，就连比邻的澳门也凭神像雕刻工艺获得2008年国家级非遗之「奖项」。 有人说非遗不过是人有我有，纯粹锦上添花；也有人说，中国在维护主权和领土完整的概念下，又怎能在文化层面少了香港一席？香港能够「出产」一个非遗，中国在全球的文化图谱中就多一个筹码。 姑勿论背后原因为何，由于「保护非物质文化遗产公约」也适用于香港，香港特区政府就有责任找出和保护濒危失传、与社会关系密切及具香港独特性的文化传统。现在起步虽迟，但为时未晚。 「遗产」的管理问题 不过，既然政府要展开普查，另一个问题来了。民间传统应该是属于民间的，并由民间自行发展，还是属于官方，由政府承担保护与管理？ 据政府委聘负责首期普查的香港科技大学华南研究中心主任廖迪生表示，政府至今仍未有任何政策配合或承诺给予全面的保护，所以，即使清单出炉，有些遗产仍有可能难逃「破产」的命运。他强调制作非遗清单只是第一步，更重要的是如何保护这些项目。 再问民政事务局，曾德成局长除了曾向立法会议员表示，制定清单是向国家文化部申请列为国家级非遗的第一步，进而再向联合国教科文组织提出申报为世界非物质文化遗产，他所提出的，就是以遗产作招徕吸引外地游客，「以提升香港作为旅游目的地的吸引力」。 这才是令人担心的地方。「非遗」这个金漆招牌在中国许多地方都有点石成金之效。戴上这个冠冕，民俗文化很容易沦为生财工具、游客的消费品，连婚嫁仪式也可用来表演，完全违背了保护非遗的原意。曾德成之言，是否意味着香港也要跟着祖国一起走上同一条路？
英文翻译价格
英文翻译价格 根据以英文作为母语的人数计算，英文是最多国家使用的官方语言，英语也是世界上最广泛的第二语言，也是欧盟，最多国际组织和英联邦国家的官方语言之一。但仅拥有世界第二位的母语使用者，少于标准汉语。上两个世纪英国和美国在文化、经济、军事、政治和科学上的领先地位使得英语成为一种国际语言。如今，许多国际场合都使用英语做为沟通媒介。英语也是与电脑联系最密切的语言，大多数编程语言都与英语有联系，而且随着网络的使用，使英文的使用更普及。英语是联合国的工作语言之一。 为了方便大家了解英文翻译价格，小编在目前汇集最多翻译团队的高校译云上面获得了不同翻译精英团队所展示的价格。 暨南大学翻译中心：中英---普稿---150---千字英中---普稿---250---千字 武汉理工大学-外国语学院MTI翻译中心 ：中英互译中英130-150 英中100-130 华中科技大学-翻译研究中心 ：中英互译中英120-150 英中100-120 湖南科技大学MTI中心：中英---普稿---150---千字 上海师大外国语学院翻译中心： 中英---普稿---200元---千字英语普通文本译成汉语---120元---千字西南大学翻译中心：中英---普稿----300---千字英中---普稿---200---千字 上海理工大学MTI翻译中心：中英---普稿---100---千字 南京财经大学外国语学院翻译研究中心：中英---普稿---100---千字 一般英文翻译价格是是在100—300元每千字，根据译员质量、翻译内容、需要的时间等都会有一定的波动，所以以上价格供大家参考，具体的可以准备好稿件了去问，这样会更加准确一些。
翻译服务收费标准
翻译服务收费标准 一、笔译人民币元/千字中文( 加急加收30% —70% ，专业加收50% ) 语种中译外外译中外译外 英语170 140 面议 日语170 140 韩语190 160 德语220 180 俄语220 180 法语220 180 意大利语280 250 西班牙语280 250 葡萄牙语290 260 阿拉伯语350 320 越南语430 400 荷兰语510 460 波兰语380-480 360-40 塞尔维亚语370-470 420-530 泰国语260-380 280-520 老挝语320-420 370-480 印度语320-420 370-480 希腊语370-470 420-530 哈萨克语280-380 300-410 瑞典语300-400 340-450 丹麦语320-420 370-470 印度尼西亚语330-450 350-460 蒙古语300-400 350-460 1、字数计算：以中文版稿件在Windows word文档显示的字符数（不计空格）为基准。也即包含了标点符号，因其为理解语义的必需。 2、图表计算：图表按每个A4页面，按页酌情计收排版费用。 3、外文互译：按照中文换算，即每个拉丁单词乘以二等于相应的中文字数。 4、日翻译量：正常翻译量3000-5000字/日/人，超过正常翻译量按专业难易受20%加急费． 5、付款方式：按预算总价的20%收取定金，按译后准确字数计总价并交稿付款。 6、注意事项：出差在原价格上增加20%，客户负责翻译的交通、食宿和安全费用。 二、口译价格： (1) 交传报价（元/人/天，加小时按100-150元/小时加收费用）类型英语德、日、法、俄、韩小语种 一般活动700 800 1500 商务活动500-1200 500-1500 800-3000 中小型会议1200-3000 1500-3000 2500-3000 大型会议1200-4000 2500-6000 4000-9000 (2) 同传报价（元/人/天） 类别中-英互译日、韩、德、俄、法、韩-中互译小语种-中互译 商务会议5000-8000 6000-10000 8000-10000 中小型会议5500-8000 7000-12000 8000-12000 大型国际会议6000-9000 8000-12000 12000-16000
浙江杭州小语种翻译公司报价
加快实施“走出去”战略是适应全球化新形势和我国发展新变化，培育参与和引领国际合作和竞争新优势的重要举措。据预测，今后五年，我国对外投资规模总量超过5000亿美元，并且年增速在10%以上。浙江是外贸大省，境外经贸合作区建设走在全国前列，境外投资一直保持高速增长，《浙江省利用外资和境外投资“十三五”规划》也作出了明确部署。跨境法商论坛拟计划在杭州举办，将促进浙企与马来西亚、新加坡相关机构的直接对话，帮助浙江企业进一步拓展“一带一路”市场，寻找国际合作渠道，促进有效投资。 各式各样大手笔的贸易投资合作，也预示着新的发展机会。对从事语言服务工作的人士无疑是好消息。据中青在线记者报道，在之前“一带一路”国际合作高峰论坛会议中，现场的同传耳机里，18种工作语言分别是：汉语、英语、法语、俄语、西班牙语、柬埔寨语、捷克语、匈牙利语、印度尼西亚语、哈萨克语、老挝语、蒙古语、波兰
语、塞尔维亚语、土耳其语、越南语、日语和韩语。每个座位上都放有一张列举了这些语言的名单，除了标注与同传耳机频道对应的16种语言以外，同传耳机中的第17、第18频道分别是日语和韩语。 这无疑是翻译从业者的一剂强心针，不少小语种从业者和学习者表示，政府、学校、企业等对于小语种的关注度，现在越来越高了。更日常的商务沟通、贸易合作等等势必越来越密集和频繁，毫无疑问，国内的翻译人才供不应求，高质量的翻译人群，已经成为最抢手的人才。 随着翻译需求的不断加大，国内的翻译产业开始蓬勃发展。从2012年到2015年，我国翻译专业硕士(MTI)学位授予点也由2007年的15所大学增加至2016年的215所大学，国内翻译公司数量已达到72，485家。咱们先了解一下市场小语种的翻译报价参考： *以上报价为人民币基准价，不含税金，仅供参考，具体报价根据需译资料数量、领域、难易等具体确定！
中译英翻译报价单中译英翻译价格
精诚翻译公司五周年五折优惠中5年经验！先翻译后付费学生客户送50元优惠券，可以搜索精诚翻译公司者50元翻译找到我们 The book of“Chinese Language" aims at improving students' Chinese accomplishment and emphasizes on humanity, instead of literature or humanism. The book mainly consists of literary works, and also involves philosophy, history, art, science and technology, so as to interest students of different majors and help broaden their vision. 1Ancient literature 2 Modern literature 3 English-Chinese translation In the textbook of Public English, one third of contents have been updated, therefore, the contents become more interesting, comprehensive and practical. The attached vocabularies enable students to master the law of learning vocabularies, and the actual use of English sentences help improve their ability of using English. "Fundamentals of Management" is a subject which discusses basic theories, principles, management functions and general method of management activities, it also combines theories with practice closely. Through studying this course, students should be familiar with basic concept of management, relevant knowledge system and thoughts by various management school; understanding basic management theories and relevant principles in depth, as well as planning, decision-making, organization, leadership, motivation, control, communication and other basic management functions and methods. "The History of Chinese Culture" The book' author is an contemporary Chinese culture expert who has published many cultural works. This book focuses on evolution of academic thought and culture, at the same time, it also consider behavior, system,
山东青岛小语种翻译公司报价
“一带一路”倡议提出五年来，青岛市紧紧围绕“新亚欧大陆桥经济走廊主要节点城市”和“海上合作战略支点”功能定位，积极融入大局，深入贯彻国家政策，在经贸合作、外事外宣、金融保障、海洋科技合作等领域着力打造对外开放新高地。对“一带一路”沿线国家累计投资69.8亿美元，承包工程完成营业额130.6亿美元。全市银行机构累计为“一带一路”沿线国家和地区有投资项目的84家大型企业授信860.3亿元，贷款余额445.1亿元，支持3000余家企业与“一带一路”沿线61个国家开展贸易投资与建设合作。市级领导出访沿线国家达50余批次；接待国外重要来宾、团组来访530余批次，友城总数达到70个，遍及全球37个国家。 各式各样大手笔的贸易投资合作，也预示着新的发展机会。对从事语言服务工作的人士无疑是好消息。据中青在线记者报道，在之前“一带一路”国际合作高峰论坛会议中，现场的同传耳机里，18种工作语言分别是：汉语、
英语、法语、俄语、西班牙语、柬埔寨语、捷克语、匈牙利语、印度尼西亚语、哈萨克语、老挝语、蒙古语、波兰语、塞尔维亚语、土耳其语、越南语、日语和韩语。每个座位上都放有一张列举了这些语言的名单，除了标注与同传耳机频道对应的16种语言以外，同传耳机中的第17、第18频道分别是日语和韩语。 这无疑是翻译从业者的一剂强心针，不少小语种从业者和学习者表示，政府、学校、企业等对于小语种的关注度，现在越来越高了。更日常的商务沟通、贸易合作等等势必越来越密集和频繁，毫无疑问，国内的翻译人才供不应求，高质量的翻译人群，已经成为最抢手的人才。 随着翻译需求的不断加大，国内的翻译产业开始蓬勃发展。从2012年到2015年，我国翻译专业硕士(MTI)学位授予点也由2007年的15所大学增加至2016年的215所大学，国内翻译公司数量已达到72，485家。咱们先了解一下市场小语种的翻译报价参考： *以上报价为人民币基准价，不含税金，仅供参考，具体报价根据需译资料数量、领域、难易等具体确定！ 尽管翻译人才输出与翻译机构成立都发展迅速，但真正能满足企业发展需求的高质量翻译机构却并不好找。一方面是因为从事翻译行业的，不少都是翻译新手，没有行业经验和过硬实力，另一方面，也是因为翻译机构资质参差不齐，没有统一的规范，翻译价格和翻译质量也受到影响。怎么选择靠谱的翻译机构就成为企业的困惑点。
小语种翻译现状
小语种翻译现状 目前很多大城市会出现一种奇怪的现象，当小众语种国家人代表在我国开会发言演讲时，台下的大家只能眼巴巴的望着他却不知所云，居然没人能翻译，这种现象引起了各相关部门的广泛关注。翻译达人表示说，我国很多二三线城市同声传译人才缺乏，翻译机构规模档次不够。面对二三线城市建设步伐的加快以及国际性会议的增多，高端翻译市场亟待提升档次。 国际会议尴尬 西安高新区工作的张先生讲述到，某次会议当天，来自亚洲的一些国家的部门代表轮流发言，参与会议的也都是来自周边国家的企业以及政府代表。每人配备同声传译设备，并且有同传人员现场翻译，一切看上去井井有条。可当轮到格鲁吉亚一位代表发言时，同传人员却“撤退”了。说着格鲁吉亚语的这位代表只好自顾自说了半天，下面参会人员看上去一片茫然。 人才严重匮乏 目前大多二三线城市的小语种翻译人员都是从北京、上海、广州这些大城市请过来的，并且价格不菲！同声传译人才对翻译水平的要求最高，不仅要具备良好的口语功底，还要对当地的文化有所了解，一般都需要有国外生活几年的经历才能胜任。大多二三线城中外语翻译人才还仅仅停留在英语方面，小语种翻译人才严重匮乏。而能提供大多小语种翻译的，一般只有像翻译达人这样的翻译平台。 小语种翻译专业开设少 导致小语种翻译人才稀缺的因素有很多，其中有一个因素就是开设小语种专业的高校少。比如小语种罗马尼亚语，全国仅有1所大学开设了这个专业，即北京外国语大学。还有上文提到的格鲁吉亚语，目前在我国还没有一所大学开设了
这个专业。曾经北大给俄语专业的学生开设了这个课程，请的格鲁吉亚的外教讲授的，但是因为外教的变动，这课程现在也没有了。因为开设小语种的学校少，学习该语种的人自然也少。随之就是翻译的价格水涨船高。可以说，翻译人才越难找，收费也就越贵。 翻译市场亟待壮大 人才的匮乏会明显的限制整个翻译市场的壮大与发展，国际性会议的大幅度增多，更导致了小语种人才严重稀缺。而懂得小语种的翻译人才，目前正在成为各大公司热捧的对象。甚至因为人才稀缺，企业在招聘标准上也低出不少，如某些公司招聘负责商务谈判及日常翻译工作的俄语翻译，已经打出“应届毕业生也可”的条件。所以，目前我国的小语种翻译市场亟待扩大，也需要更多的小语种翻译人才。