共查询到20条相似文献，搜索用时 62 毫秒
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《科学中国人》2015,(26)
目的构建SLO原核表达质粒,重组蛋白的诱导表达并纯化。方法提取链霉菌溶血素O模板DNA,PCR法扩增slo基因。构建融合表达重组质粒p GEX-6p-1-slo和pet32a-tev-slo,将正确的重组质粒转化至大肠杆菌BL21和大肠杆菌BL21-DE3,使用异丙基硫代βD半乳糖苷(IPTG)诱导表达重组融合蛋白。采用亲和层析纯化重组蛋白,切除标签后,再通过亲和层析纯化,获取SLO重组蛋白。结果 PCR扩增出slo基因,基因片段(1700 bp)与理论一致。经SDSPAGE,蛋白质印迹显示重组蛋白相对分子质量为55 k Da,与数据库中的相对分子质量结果相符。结论成功构建了slo原核表达质粒,表达并纯化了SLO重组蛋白。 相似文献
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《科学中国人》2015,(26)
目的:构建Visfatin原核表达质粒,诱导表达并纯化重组人内脂素(Visfatin)。方法:从人LO2细胞中提取总RNA后,经RT-PCR法得到人Visfatin的c DNA序列,用以构建带HIS标签的Visfatin重组质粒。然后将重组质粒转化到大肠杆菌BL21中,用异丙基硫代-β-D-半乳糖苷(IPTG)诱导表达,用Ni柱亲和层析纯化融合蛋白。结果:PCR扩增出的目的基因片段为1600 bp左右,与理论大小一致。SDS-PAGE、Western blot、质谱分析数据证明了蛋白是Visfatin融合蛋白。结论:成功的表达并纯化得到带HIS标签的Visfatin融合蛋白。 相似文献
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本试验用VP1基因C末端与表达载体连接,转化到大肠杆菌中,经IPTG诱导表达,收集不同时间段的菌液进行SDS-PAGE,结果表明重组的VP1基因C末端蛋白在大肠杆菌中能高效表达,表达的目的蛋白分子量约为16kd,而且重组菌诱导裂解物可在16kd处看到一条特异的蛋白条带,与预期大小一致,阴性对照未出现表达条带,而且重组菌在IPTG终浓度为1mmoL/L诱导6 h时表达量最大。表达产物主要以包涵体的形式存在,提取和裂解包涵体后,进一步纯化表达的目的蛋白,可获得纯度达70%以上的目的蛋白。 相似文献
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在成功建立稳定分泌鼠抗人B7-2杂交瘤细胞株基础上,扩增并克隆出该单克隆抗体的重链(VH)和轻链(VL)可变区基因。通过重叠延伸PCR(SOE-PCR)方法,在VH和VL可变区基因之间引入连接肽(Gly4Ser)3,体外构建抗人B7-2单链抗体(B7-2 ScFv)基因。为便于表达产物的纯化,在抗人B7-2 ScFv的C端增加了6×His tag序列。将其克隆至表达载体pET32a并在大肠杆菌中进行原核表达。SDS-PAGE和Western-blot分析结果表明,抗人B7-2 ScFv在大肠杆菌BL21(DE3)菌中获得表达,重组融合蛋白的相对分子量约为43 kD,表达产物以不溶性包涵体形式存在,经溶解包涵体,体外复性和Ni-NTA亲和柱纯化,获得了高纯度的抗人B7-2 ScFv,纯化蛋白得率约16.2%。成果为抗人B7-2单链抗体的生物学活性研究和抗肿瘤药物开发奠定了基础。 相似文献
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溶藻弧菌铁调蛋白基因的克隆、表达及其特征分析 总被引：1，自引：0，他引：1
铁为一切有机体所必需。对铁摄取与利用的调控在细菌生长和铁毒性中起重要作用。在革兰氏阴性细菌中,铁摄取调节蛋白(Fur)主管铁的代谢。溶藻弧菌是人和海洋动物共感染的一种主要病原弧菌。本研究克隆表达了溶藻弧菌的fur基因,并分析了其相关特征。该基因序列全长994 bp,其中包含450 bp的开放读码框,编码150个氨基酸残基的蛋白,推导的相对分子量为16.78 kD.在编码序列的上下游,RBS序列-、10序列-、35序列和二个潜在的转录终止子分别得到确定,但没有发现存在于大肠杆菌fur基因中的"铁盒子"。氨基酸序列的中段从G46位至D104位,为高度保守区域,其中包括含组氨酸丰富铁结合模型(H3-D-H-L-V-C-L-D-C-G)。SDS-PAGE分析表明,fur基因可在大肠杆菌中大量表达,带His-Tag的重组融和蛋白经镍亲和层析得到纯化,Western-blot分析结果显示,Fur蛋白具有免疫反应性。为进一步研究Fur蛋白的结构与功能打下基础。 相似文献
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目的:扩增弓形虫SAG2基因,构建pGEX-4T-1-SAG2重组质粒,在大肠杆菌中表达。方法:设计并合成引物,经PCR法扩增SAG2基因序列,与pMD18-T载体连接,构建pMD18-T-SAG2质粒,经酶切鉴定、测序,与pGEX-4T-1载体连接,构建pGEX-4T-1-SAG2质粒。将重组质粒转入大肠杆菌诱导表达,进行SDS-PAGE和Western-blot鉴定。结果:扩增出约561bp的SAG2基因序列,成功构建pGEX-4T-1-SAG2质粒,并在大肠杆菌中得到高效表达,融合蛋白的表达量占菌体总蛋白的40%。SDS-PAGE电泳显示pGEX-4T-1-SAG2的融合蛋白的分子量约为45kd。Western-blot显示融合蛋白有良好的抗原性。结论表达质粒在大肠杆菌中得到高效表达,为进一步研究弓形虫病的诊断做好准备。 相似文献
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《中国科学院院刊(英文版)》2008,22(1):2-3
CAS should stick to the principle of rendering service to, and giving impetus for, the development of China＇s science enterprise by making S＆T innovations, said CAS President LU Yongxiang. The CAS president made the remarks in a recent talk to communicate the gist of the winter session of the Party＇s Leading-member Group at CAS, which was held from 7 to 11 January in Beijing. 相似文献
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《中国科学院院刊(英文版)》2008,22(1)
With great care, Dr. ZHOU Zhonghe takes out a package wrapped by cotton tissue from a drawer and says: "This is the gem of our collections: the fossil of a bird that lived 125 million years ago!" Then, pointing at a tiny mound, he explains: "Look, this is the claw and that is the head. It was in the egg shell and ready to hatch ... The species fell into a family of waterside inhabitants." 相似文献
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Prof. SUN Changpu from the CAS Institute of Theoretical Physics coworkers from University of Basel in Switzerland have worked out a way - at least in theory - to split a beam of molecules according to their chirality. The technique involves passing the molecules through three different laser beams is similar to the famous Stern-Gerlach effect whereby a beam of atoms passing through a magnetic field is split in two according to the atoms' spin states 《中国科学院院刊(英文版)》2008,22(1):23-23
Prof. SUN Changpu from the CAS Institute of Theoretical Physics and coworkers from University of Basel in Switzerland have worked out a way --at least in theory --to split a beam of molecules according to their chirality. The technique involves passing the molecules through three different laser beams and is similar to the famous Stern-Gerlach effect, whereby a beam of atoms passing through a magnetic field is split in two according to the atoms＇ spin states （Phys. Rev. Lett. 99 130403）. 相似文献
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《中国科学院院刊(英文版)》2014,(3):212-213
<正>In February 2014,the 30th Chinese Antarctic inland expedition team reported that they have successfully conducted the seeing measurement at the Antarctic Taishan Station using a Differential Image Motion Monitor(DIMM)developed by the Nanjing Institute of Astronomical Optic and Technology(NIAOT),Chinese Academy of Sciences.In astronomy,seeing is a physical parameter that describes the irregular movement and blur of the star 相似文献
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XIN Ling 《中国科学院院刊(英文版)》2014,(4):270-271
Amassive overhaul is coming to the Chinese Academy of Sciences （CAS）. To straighten its ＂fragmented, inefficient research＂, the academy has decided to restructure its 104 institutes for better teamwork. 相似文献
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SONG Jianlan 《中国科学院院刊(英文版)》2014,(4)
<正>DNA demethylation in mammalian zygotes,a biochemical process through which the parental genomes let go the epigenetic marks from their"past lives"to obtain totipotency,has long fascinated biologists:how does it happen?A joint team at the CAS Shanghai Institutes for Biological Sciences tells a surprising story. 相似文献
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China is one of the fastest growing economies worldwide. Are science and research also in the fast lane? We talked with LUO Yuan, scientist at the Institute of Coal Chemistry in Shanxi, and Axel Mosig, Professor at Ruhr University in Bochum, who has first-hand experience working and living in China. 相似文献
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Joachim Trümper 《中国科学院院刊(英文版)》2014,(2):201-201
<正>"In 1979 two Chinese scientists joined our institute in the framework of the historical CAS-MPG agreement.This marked the beginning of a longstanding fruitful collaboration.Many visitors followed and contributed to our scientific programs.–My first visit to China took place in 1981–with stays at CAS institutes in Bejing,Nanjing,Shanghai and Kunming,where I received unforgettable impressions of 相似文献
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《中国科学院院刊(英文版)》2014,(3)
<正>May 15,2014Publicly funded scientific research is an important means for the society to create knowledge,support innovation,and promote growth.Research articles from such research are knowledge resources to be shared by all.Their open access will facilitate the dissemination and utilization of knowledge,turn the knowledge produced by public investment effectively into innovation and development capabilities of the society,ensure that public knowledge benefi ts all,and strengthen the innovation driven development strategy.Therefore,Chinese Academy of Sciences(CAS)issues at the present stage the following open access policy for articles from publicly funded scientifi c research projects:CAS requires its researchers and graduate students to 相似文献
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《中国科学院院刊(英文版)》2014,(3)
<正>Between February and March of 2013,the source of avian-origin H7N9 human infection was discovered in Shanghai and Anhui,China.Prior to this,H7N9 has only exhibited characteristics of a low pathogenic avian infl uenza virus,carried by chickens with no known cross-species transmission.By the end of August 2013,Chinese mainland reported a total of 134 confi rmed cases of H7N9 human infections across12 provinces and 42 cities,resulting in 45 deaths.In October and November,Zhejiang Province found three new cases and Guangdong found one case of human infection,suggesting the possibility of a comeback in the fall and winter seasons.Researchers led by Prof.GAO Fu(George Fu Gao)at the Institute of Microbiology,CAS and Beijing Institutes of Life Science,have made progress on understanding 相似文献