表達克隆化 的英文怎麼說
中文拼音 [biǎodákèlōnghuà]
表達克隆化
英文
expression cloning phenotype cloning- 表 : Ⅰ名詞1 (外面;外表) outside; surface; external 2 (中表親戚) the relationship between the child...
- 達 : Ⅰ動詞1 (暢通) extend 2 (達到) reach; attain; amount to 3 (通曉; 明白) understand thoroughly...
- 克 : 克i 動詞1 (能) can; be able to 2 (克服; 克制) restrain; control 3 (攻下據點; 戰勝) overcome...
- 隆 : 隆Ⅰ形容詞1 (盛大) grand2 (興盛) prosperous; flourishing; thriving 3 (深厚; 程度深) deep; in...
- 達克 : d'ak taan
- 隆化 : longhua
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Screening target ' gene of artemisinin antimalarials using drug - western from cdna expressing library : 12 b - deoxoartemisinyl - ( 4 ' - oxyacetic acid ) phenyl ether was linked to bsa by using of edc cross ! inker and the product acted as drug - probe
對重組pmd一18一t克隆載體及pqe一30表達載體雙酶切,提取tctp基因和pqe一30空載體並使二者重組,然後轉化m15 ,挑取陽Construction, expression and purification of his - hmgb1 and novel antigen in autoimmune hepatitis
1的克隆表達鑒定及蛋白純化Degenerate oligonucleotides to highly conserved regions of cucumis melo 1 - aminocyclopropane - 1 - carboxylic acid ( acc ) oxidase gene were used to prime the amplification of fragment of 128bp by ploymerase chain reaction ( pcr ) in samples of genomic dna from fruit of cucumis melo l. cv hetao flesh, which was cloned into plasmid vector pmd - 18 - t. the clon of antisense orientation were selected, and it was inserted downstream of camv35s promoter and enhancer " " of tmv into the plant expression vector pbinyxw, antisence expression vector pbinya was constructed. at the base that pollination and fertilization of cucumis melo l. cv hetao was studied, using pollen tube pathway transformate cucumis melo l. cv hetao, 76 fruit had been obtained, moreover, hardness and content of sugar were analysed
本實驗以河套蜜瓜果肉基因組dna為模板,用甜瓜acc氧化酶基因特異寡核苷酸鏈為引物進行pcr擴增,得到128bp的擴增產物。將得到的擴增產物克隆到質粒載體pmd - 18 - t上,篩選反向克隆,然後將其反向構建到植物表達載體pbinyxw的camv35s啟動子和tmv增強子「 」的下游,構建成反義表達載體pbinya 。並在對河套蜜瓜授粉受精生物學研究的基礎上,通過花粉管通道法轉化河套蜜瓜,共獲76顆瓜,並進行了硬度和含糖量的分析。Confocal microscopy observation followed immune - fluorescence staining with anti - gp130 showed that gp130 could interact with tle1 at the cytomembrane region. moreover, this interaction inhibited the concentration of tle1 into nucleus
為了進一步證實上述發現,我們表達並純化了gst - gp130胞漿區融合蛋白和gst - tle1獨特性片段融合蛋白,並制備了特異性抗tle1多克隆抗體。In this paper, first strand cdna of 3abc gene was synthesized using template rna extracted from cells infected with fmdv. the complete 3abc gene about isoobp was amplified by pcr and ligated into pgem - t easy vector. after transforming e. coli dh5 a, ampicillin resistant colonies were isolated and plasmid dna was prepared and analyzed by restriction analysis and pcr. presence of the full length 3abc gene was verified by nucleotide sequence analysis and the plasmid containing the expected sequence was named as pgem - 3abc. comparing the aquired sequence of 3abc with that of reference strains, the homology is more than 99 percent. the pgem - 3abc was digested with sal i and bgl ii and ligated into xho i and bgl ii - digested expression vector ptriex - 4 neo. lt was identified by restriction analysis and pcr and sequencing that this fragment had a 17bp deletion hi the nucleotide sequence 708bp of 3abc gene, which happened to form a terminator codon behind 3ab gene, but it contained the complete open reading frame ( orf ) of 3ab gene. positive clones were selected and induced with lmmol / l isopropyl - d - galactoside ( iptg ), bacteria were detected by sds - page and western blotting after properly treated. the results showed that the 3ab gene expressed successfully in e. coli and 33. 5ku fusion protein can be recognized by the positive bovine serum of fmdv. the amount of target protein is over 26 % of the total bacteria protein by gel thin layer scanning analysis
擴增產物連接到pgem - teasy載體中,轉化大腸桿菌dh5菌株,篩選氨芐青霉素抗性菌落,提取質粒經酶切鑒定、 pcr分析以及確證性測序證明,所克隆的1500bp左右的片段含有完整的3abc基因,與國外參考序列相比,同源性在99以上。將重組質粒pgem - 3abc和表達載體ptriex - 4neo分別用sal和bgl與xho和bgl消化后,亞克隆3abc基因至原核表達載體ptriex - 4neo中,通過酶切鑒定、 pcr擴增以及序列分析,發現克隆到ptriex - 4neo載體上的片段於3abc基因708bp處出現了17bp的缺失,碰巧在3ab基因后形成一終止密碼子,但3ab基因的閱讀框架完整,選出含有3ab基因完整閱讀框架的陽性克隆,用iptg誘導表達,收集菌液進行sds - page電泳、 westernblotting分析,結果表明, 3ab基因在大腸桿菌中成功表達,其表達產物為分子量33 . 5ku的融合蛋白,並能被口蹄疫病毒陽性血清識別。經薄層掃描分析,表達量占總蛋白量的26以上。Many studies show that leafy is high homolog even among distantly related plant species. exception of these, little studies on tissue culture and transformation of ginkgo have been done. this paper emphasizes on the isolation, cloning and analysing two ginkgo orthologs of leafy from the male tree
為此,本實驗從銀杏leafy同源基因的克隆入手,分析其雌雄株lfy基因結構差異,構建lfy基因的植物正義反義表達載體,建立矮牽牛遺傳轉化體系,以研究銀杏lfy同源基因的功能,同時建立了銀杏組織培養體系,為銀杏的遺傳轉化和提早開花結果奠定基礎。Porcine transmissible gastroenteristis is an importan contagious disease endangering the development of swine. in other to establish a rapid diagnosis method and provide effective immunogenic products, the nucleoprotein ( n ) gene of porcine transmissible gastroenteristis virus ( tgev ) was cloned. expressed and its expressed product was purified
為建立對豬傳染性胃腸炎快速有效的診斷方法,並試圖在預防上提供有效的免疫制劑,本論文首次在我國對豬傳染性胃腸炎病毒核衣殼蛋白基因進行了克隆、鑒定、表達及重組核蛋白的純化;並在細胞上對重組核衣殼蛋白抗體的中和效力進行了測定。It is interesting that pma plus calcium ionophore a23187 can inhibit pma - induced pta1 expression, and this effect ca n ' t be reversed by calcmeurin inhibiter fk506. pta1 mabs can inhibit ctl activation and differentiation in mixed lymphocyte culture system when added at the beginning of the culture but can induce platelet activation and aggregation in the fc dependent manner. in 1997, pta1 cdna was cloned from cdna library of tpa activated jurkat cells, which belongs to immunoglobulin superfamily ( igsf ) with two v - like domains of extracelluar region of pta1
Il - 2 、 tnf - 、 pma可以使t細胞pta1表達上調, tgf -可以下調pta1的表達,而pma加上鈣離子載體a23187可以顯著抑制pma的上調作用,且這種抑制作用不被calcineurin抑制劑fk506所逆轉, 1997年burns教授從pma活化的jurkat細胞cdna文庫中克隆了pta1cdna全長,證實pta1是一個新分子,屬于免疫球蛋白超家族,胞膜外區有兩個v樣結構域。Marek " s disease virus ( mdv ) phosphoprotein 24 ( pp24 ) gene was amplified from md11 strain by polymerase chain reaction ( pcr ). then we cloned it into the downstream of gst gene according to the right open reading frame ( orf ) in pgex - 6p - l vector
本研究將型mdvmd11株的pp24基因的完整orf克隆入原核表達載體pgex - 6p - 1中,重組質粒pgex - pp24轉化bl21宿主菌后,經iptg誘導表達。The recombinant expression plasmids prt - p450nor and pet - p450nor were constructed by inserting p450nor gene into the bamh i / hind iii site of the prokaryotic expression vector prset and pet28, then they were transformed into e. coli bl21
本研究將已克隆的真菌細胞色素p450nor基因插入原核表達質粒載體prset和pet28的bamhi / hind位點,成功構建重組表達質粒prt - p450nor和pet - p450nor ,並轉化到e . colibl21 。As a popular vegetable, tomato is rich in vatamin a, b, p and other nutrients ; among them, lyxopene can prevent prostate gland tumor. so introducing stilbene synthase gene into tomato to get new health - protection tomato will have important social and economic effect. our research is made up of four parts ; the cloning and sequencing of stilbene synthase ; tomato genetic transforming mediated by agrobacterium - lba4404 ; quantification of resveratrol in transgenic tomato by hplc analysis
本研究包括四個方面的內容:芪合酶基因( stilbenesynthasegene )的克隆與全序列分析;芪合酶基因( stilbenesynthasegene )植物表達載體的構建;農桿菌lba4404介導的番茄遺傳轉化: hplc技術分析轉基因植株中表達白藜蘆醇的含量。Plants are thought to remove na + from the cytoplasm by transporting it into the vacuolar or out of the cell using na + / h + exchangers localized in the vacuolar and plasma membranes, respectively. sos1 encoding a plasma membrane na + / h + antiporter and atnhxl encoding a vacuolar na + / h + antiporter were isolated from glycophytic arabidopsis thaliana, and overexpression of atnhxl and sos1 in arabidopsis thaliana increased the salt tolerance of transgenic plants significantly
目前,擬南芥細胞內控制na ~ +外排的基因sos1及離子區隔化基因atnhx1均已克隆, sos1及atnhx1在擬南芥中的過量表達顯著提高了轉基因植株的耐鹽性,開創了降低na ~ +毒害的基因操作新途徑。Green fluorescent protein ( gfp ) gene was conjugated to the 3 " end of the pap gene in order to screen easily of the transgenic cotton plants. the combined gene was cloned into plant expression vector pbi121 and then transformed. about 5000 seeds of the transgenic cotton were obtained and the some seedlings of the transgenic cotton could give a bright green fluorescence in the dark condition when the cotton seedlings were irradiated with ultraviolet rays
為了便於轉基因棉花後代的篩選,在pap基因的3 』端融入了綠色熒光蛋白gfp )基因,然後將融合基因克隆在植物表達載體pbi121上,再進行遺傳轉化,得轉基因棉花種子5000餘粒,將種子播種長到于葉展開時,先在黑暗中用紫外燈照射,查找表現綠色熒光的幼苗,然後再用地高辛( dig )標記的pap基因特異性探針對這些棉花進行點雜交,最後發現有8株棉花表現陽性反應,說明pap基因的確己經轉到了棉花的基因組中,其棉花黃萎病的抗性鑒定正在進行之中。Abstract : plant responses to salt stress via a complex mechanism, including sensing and transducing the stress signal, activating the transcription factors and the corresponding metabolizing genes. since the whole mechanism is still unclear, this review emphasize the biochemical events during the plant adaptation to salt stress referring to an index of importance : the homeostasis in cytoplasm, the biosynthesis of osmolytes and the transport of water. most of these biochemical events were elucidated by study of halophyte and salt - sensitive mutations, also many important genes involved were cloned and used to generate stress - tolerance phenotypes in transgenic plants. on the other hand, about the molecular mechanism in signal transduction, the research of arabidopsis mutations and yeast functional complementation provided helpful traces but not full pathway
摘要植物對鹽脅迫的耐受反應是個復雜的過程,在分子水平上它包括對外界鹽信號的感應和傳遞,特異轉錄因子的激活和下游控制生理生化應答的效應基因的表達.在生化應答中,本文著重討論負責維持和重建離子平衡的膜轉運蛋白、滲調劑的生物合成和功能及水分控制.這些生理生化應答最終使得液泡中離子濃度升高和滲調劑在胞質中積累.近年來,通過對各種鹽生植物或鹽敏感突變株的研究,闡明了許多鹽應答的離子轉運途徑、水通道和物種特異的滲調劑代謝途徑,克隆了其相關基因並能在轉基因淡水植物中產生耐鹽表型;另一方面,在擬南芥突變體及利用酵母鹽敏感突變株功能互補篩選得到一些編碼信號傳遞蛋白的基因,這些都有助於闡明植物鹽脅迫應答的分子機制。After sds - page and densitometric scan analysis, the expression level of hng fusion protein is above 40 % and m - insulin fusion protein above 50 %. western - blot result demonstrated m - insulin fusion protein had specific reaction with mouse anti human insulin antibody, we got hng fusion protein and m - insulin fusion protein with purity of above 80 %
今士考個二目卜乙成功構建了ptxbi一hng及ptxbi一m一insulin原核表達克隆,並獲得了高效表達,經過純化得到純度人於80 %的融合蛋白,並對人胰島素突變體融合蛋白進行了初步活性測定。Purpose 1 construction of prokaryotic high expression vector of human platelet factor 4 ( h pf4 ) 2 expression and purification of r h pf4 3 bioassay of r h pf4 methods according to the modulation character of eukaryotic protein expression in prokaryotic cells, we design a pair of particular primers, and construct a prokaryotic expression vector pbv220 - r hpf4 by dna polymerase chain reaction ( pcr ) and dna recombinant technic. the expression plasmid was identified with pcr and dna sequencing. pbv220 - r hpf4 was transformed into e. coli dh5a, bl21 ( de3 ) and induced by increasing the temperature to 42. we identified the expression protein by sds - page and western - blotting
目的1人血小板因子4 ( hpf4 )原核高效表達克隆的構建2重組hpf4的表達及分離、純化工藝研究3重組hpf4的特性研究方法根據原核細胞表達真核蛋白的基因表達調控特點,設計合成一對特異引物,在pt7 - 7 - rpf4表達質粒的基礎上,應用聚合酶鏈式反應( pcr )對其cdna進行改造,通過dna重組技術構建成重組hpf4原核表達質粒pbv220 - rhpf4 ,用快速pcr檢測法、 dna測序分析,鑒定重組hpf4表達質粒的正確性。Western - blotting result demostrated rhpf4 had specific reaction with rabbit anti - hpf4 antibody. our system improve the expression level of r hpf4 by 80 fold compared with pt7 - 7 - r hpf4. after purified and renatured, r hpf4 prepared by our methods has bioactivity like wide hpf4. our study establish a stable base for further reseach of the h pf4 and provide a theoretics gist for modulative mechanism of eukaryotic protein expression in prokaryotic cells
我們構建的rhpn原核高效表達系統經m page及凝膠密度掃描分析結果表明, rhpf4表達量占菌體總蛋白量的25 30 ,較原表達克隆pt7 7 rhpf4提高了近80倍,經快速高效的包涵體分高純化工藝和復性工藝, rhpf4具有野生蛋白活性。The plasmids pci - mbl54 containing full length of mutations mbl cdna were propagated hi escherichia coli xl - 1 blue, then the extracted and purified pci - mbl54 were used to transfect dhfr ( - ) chinese - hamster ovary ( cho ) cells. after screeened with g418 and cloned, 4 g418 - resistent clones were randomly selected for detection of mrna expression by rt - pcr and molecular beacons. it was found that all of the 4 positive cell clones expresse mbl analogue as detected in transcription level
抽提、鑒定、純化重組質粒后,脂質體轉染法將重組質粒導入中國倉鼠卵巢細胞( cho - dhfr ~ - )中, g418選擇轉染子並克隆化培養,經rt - pcr和分子燈塔探針雜交鑒定其mrna轉錄,獲得4株穩定表達54位密碼突變型mbl的cho細胞。These suggest the bsp proteins and its homologous proteins are ubiquitous in mammals and must have some physical significances. in our research, to find and study the important role of bsp proteinsand their related proteins in the development of zygotes and to find the new gene related to bsp, we cloned a cdna gene sequence related to bsp proteins and make it express in eukaratic cells. the fusion protein acquired are purified are used to deduce its biological functions
而在我們的實驗中為了發現和研究bsp及其相關蛋白在受精卵的發育中的重要作用,尋找bsp相關的新基因,克隆了一個與bsp蛋白相關基因的山一na序列並將該基因編碼的蛋白質在原核細胞中進行了表達和純化,將純化后的融合蛋白作了一些生物學功能方面的初步工作,發現新蛋白對tpk有一定的抑制作用,而tpk是細胞生長和代謝中十分重要的關鍵酶,這對于進一步研究新蛋白的生物學功能具有十分重要的意義。In the first part, we observed the changes of expressions of type i receptor of il - 1 in the rat and mouse brain after intraperitoneally administration of different kinds and doses antigens respectively. in the second part including two experiments we cloned rat il - 6r ' s genes by pcr, expressed them in e. coli dh5 a and cos - 7 cell, and produced il - 6r ' s polyclonal antibody which is proved having more high liter, affinity and specificity 1
第一部分採用不同種類和劑量的抗原刺激,分兩個實驗,觀察了大、小鼠腦內il - 1的i型受體表達的變化,探討了腦內參與免疫調控的核團和細胞類型;第二部分分兩個實驗,運用pcr技術克隆了大鼠il - 6r的基因並進行了原核和真核表達,制備了特異性高、親和力強和較高效價的抗il - 6r多克隆抗體,為進一步進行il - 6r的研究奠定了基礎。分享友人