質粒克隆載體 的英文怎麼說
中文拼音 [zhílìkèlōngzǎitǐ]
質粒克隆載體
英文
plasmid cloning vector- 質 : Ⅰ名詞1 (性質; 本質) nature; character; essence 2 (質量) quality 3 (物質) matter; substance;...
- 粒 : Ⅰ名 (小圓珠形或小碎塊形物) small particles; grain; granule; pellet Ⅱ量詞(用於粒狀物)
- 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...
- 載 : 載Ⅰ名詞(年) year : 一年半載 six to twelve months; six months to a year; 三年五載 three to five ...
- 體 : 體構詞成分。
- 載體 : [化學] carrier; supporter; isotopic carrier
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The total rna was isolated from pokeweed ( phytolacca americana ) leaves using the method of guanidine isothiocyante and used as template to amplify the total length and deleted mutant pokeweed antiviral protein ( pap ) gene by rt - pcr and then the pap gene was cloned into pgem - t vector. the sequencing results showed that pap gene had 99. 9 % identity comparing with the pap gene nucleotide sequence reported by lin et al ( 1991 ). the iptg - inducible expression vector containing the pap gene was constructed and transferred into e. coli bl21 ( de3 ) - plyss
將缺失型pap基因克隆到植物表達載體pbi121中,通過液氮冷凍法將重組質粒轉入農桿菌lba4404細胞中,然後採用葉盤法,在該農桿菌的介導下將pap基因導入普通煙草中,經過卡那黴素抗性篩選,最後獲得了轉pap基因的工程煙草植株,摩擦接種煙草花葉病毒( tmv ) ,與非轉基因煙草相比,能夠推遲癥狀表現達2月之久,說明pap基因能夠在其它植物體內產生有活性的高抗病毒的蛋白質。Methods : an artificial gene for fl extracellular domain cdna was synthesized by using favored genetic codons of pichia pastroris. by inserting human fl extracellular domain cdna coding 156 amino acid resi dues into pichia pastoris expression vector ppic9k containing aox1 promoter and the sequences of alpha secreting signal peptides, a recombinant expression plasmid ppic9k - fl was constructed, and integrated into the alcohol oxidase region of the host genome
為了提高外源基因的表達量,我們根據畢赤氏酵母偏愛密碼子人工合成了編碼fl胞外區156個氨基酸的cdna序列,目的序列被定向克隆到酵母分泌型表達載體ppic9k質粒上,構建ppic9k - fl表達質粒。In this study, iltv - nm98a strain and iltv - wanggang strain were multiplied in chorioallantois. a pair of primers were devised according to the nucleic acid sequence of iltv tk gene and the dna of multiplied virus was used as pattern to amplify the gene of tk by polymerase chain reaction ( pcr ). the product of pcr was linked with suitable plasmid. then, the recombined plasmid was converted to escherichia coli. the converted escherichia coli
根據已發表的iltvtk基因的核苷酸序列設計一對pcr引物,以增殖的兩株iltv的dna為模板,分別對它們的tk基因進行pcr擴增。將回收的pcr產物連接到適當的質粒載體上,轉化感受態大腸桿菌,通過篩選對iltvtk基因的陽性克隆進行擴增培養。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。The study results of this paper can serve the two scientific subjects of our teaching and research group as basic data calculation and elementary exploration. the two subjects are : constructing high activity - amylase genes by dna shuffling technology and studying on the evolution in vitro by mutational pcr ( with 5 - br - dutp as substitute partly ) and dna shuffling technology. - amylase ( ec 3. 2. 1. 1 ; 1, 4 - a - d - glucanohydrolase ) can catalyzes the hydrolysis of - 1, 4 - glycosidic bonds of starch from middle and liberates - maltose, - glucose and - limit dextrin stepwise
本試驗根據genbank已公布的黃單胞菌-澱粉酶基因的核苷酸序列由引物設計軟體premierprimer5 . 0輔助設計了一對引物( primer & primer ) ,以pbluescript ks +和puc18 / puc19質粒為載體,用常規的pcr方法從xanthomonascampestrispv . malvacearum ( smith ) dye等七株黃單胞菌( xanthomomasspp . )的基因組dna中克隆得到8個基因片段,分別命名為zhyf001 zhyf008 。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。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 result showed that the homology rate of pila gene among the 5 avian pathogenic e. coli strains tested and one human e. coli were from 89. 8 % to 91. 1 %, and the homology rate of amino acid were from 88. 5 % to 91. 8 %. the homology rate of pila gene sequence among 5 avian pathogenic e. coli strains tested and avian pathogenic e. coli reported ( serotype o1, o2, o78 ) were from 87. 8 % to 90. 2 %, and the homology rate of amino acid were from 84. 6 % to 91. 2 %. there had homology in avian pathogenic e. coli. there had some common antigen side in type 1 pili of avian pathogenic e. coli
結果表明:運用msha法檢測1型菌毛的檢出率為80 ( 36 45 ) , pcr法的檢出率為95 . 5 ( 43 45 ) , pcr方法用於1型菌毛的檢測比msha更加敏感、快速、特異性強;選擇5株優勢血清型雞源致病性大腸桿菌代表株( o _ ( 89 ) , o _ ( 119 ) , o _ ( 141 ) , o _ ( 127 ) )的1型菌毛pila基因的pcr擴增片段經純化后,分別定向克隆到puc18質粒的多克隆位點,構建了含有目的基因片段的克隆質粒,並轉化到dh5株大腸桿菌載體菌中,篩選獲得陽性克隆菌株。In this paper, a field strain of infectious bronchitis virus was isolated from proventriculus tissue, morphological observation by electron - microscope and the biological characterizations of the virus were studied, pairs of specific primers are designed and synthesized in correspondence with them, according to the published sequences of infectious bronchitis virus three structural protein ( spike protein s membrane protein m nucleocapsid protein n ) genes, the cdna of si gene, s2 gene, m gene. n gene of ib v isolate lx4 were amplified by rt - pcr and full sequences were first reported
在此基礎上,根據國內外已發表的ibv基因序列,分別設計特異性引物,應用不同引物進行反轉錄合成cdna ,分片段對ibv的主要結構基因進行pcr擴增,並分別將各個目的片段克隆到puc19載體上,在大腸桿菌dh5中實現目的基因的分子克隆,經藍白斑篩選、限制性內切酶分析、 pcr鑒定,篩選出重組陽性質粒,並對各個目的基因片段進行序列測定,從而獲得ibv主要結構基因全序列。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 。The recombinant transfer vector pbacpak - hbmp was constructed by insertion of the hbmp coding sequences into the multiple cloning site of transfer vector pbacpak. 8. bmn cell line was co - transfected with pbacpak - hbmp plasmid and linearized baculovirus bacpak6 dna by dosper agent
將克隆到的hbmp基因通過適當的酶切插入到轉移載體質粒pbac - pak8的多克隆位點中,獲得重組轉移載體質粒pbacpak - hbmp 。Since the plasmid is capable of independent replication in host cells of many dicotyledonous plants, it has been used as a cloning vector in gnetic engineering
在許多雙子葉植物中質粒在寄主細胞中是獨立的復制單元,所以可以在基因工程中用作克隆載體。In this study, the recombinant fowl - poxvirus was transfected into expressing the vp3 gene of isolated gpv h1 strain into the cef cells with fpv - 017 by liposome, which have the lacz reporter gene, earlier / latter promoters lp2ep2 of fpv, promoters p7. 5 and p7. 1 of vaccinia virus, replication unnecessary region of fpv - 017. following 6 cycles screenings, clonings, purification of blue plaques, detection of pcr and dot - elisa, which verified the genetic stable vp3 - fowlpox virus recombinant constructed successfully. this study provided the theoretical and practical foundation for development of gpv recombinant fowl - poxvirus genetic engineering vaccine, as well as provided substance preparatory for prevention the high mortality gpv
本研究採用脂質體轉染方法,將含有完整gpvh1分離株vp3基因、報告基因lacz 、禽痘病毒早晚期啟動子lp2ep2 、痘苗病毒啟動子p7 . 5 、 p11和fpv - 017復制非必須區的轉移載體質粒psy681vp3lacz與fpv - 017共轉染雞胚成纖維細胞,經6輪蝕斑克隆、篩選、表達, pcr鑒定和dot - elisa檢測,證明該重組病毒已構建成功,並獲得了遺傳性狀穩定的鵝細小病毒vp3基因的重組禽痘病毒。In order to study the function of cycling2 in vitro culturing cell line, we used pires - g2 eukaryotic expression vector transfecting human gastric cell line sgc - 7901 and human embryo kidney hek - 293 cells by lipofectamine plus reagent, and studied the function of cycling2 expression on the cell proliferation in vitro, further investigated the regulation mechanism of cycling2. at the same time, we made a study on the expression level change of cycling2 in normal gastric tissue and different type and different stage of gastric carcinoma tissue. material and method 1 material : piresneo vector was purchased from clonetech, plasmid extraction and purification kit was purchased from qiagen company ; rpmi 1640, dmem fetal calf serum were obtained from gibco / brl ; lipofectamine plus and g418 were purchased from life technologies ; ultrasensitive ? s - p kit, mouse monoclonal antibody p21wafl ( in use ), dab staining kit were purchased from maixin company
實驗材料與方法1 .實驗試劑高糖dmem 、 rpmll640和胎牛血清購自美國g山eo / brl公司; dmewf12 ( 1 : 1 )混合培養液購自美國hyclone公司;胰蛋白酶購自美國si目叮a公司; hepes由美國amersco公司分裝;脂質體轉染試劑( upofectalnineplusreageni )和以18為美國玩vitrogen公司產品; piresneo載體購自美國cloneteeh公司;質粒提取及純化試劑盒購自德國qiagen公司; ultresensitive翎s一p免疫組織化學試劑盒;鼠單克隆抗體戶3 ( do一7 )蛋白(即用型) ;鼠單克隆抗體p21waf , (即用型) ; dab染色試劑盒均購自福建邁新公司;鼠單克隆抗體pziwa曰(濃縮型) ;辣根過氧化酶標記羊抗鼠二抗購自北京中山公司; ecl試劑盒購自美國santacruze公司; dcproteinassay試劑盒購自bi 。Vp3 gene of hl isolate of goose parvovirus derived from recombinant piasmid pproex htb - vp3 was subcloned into ecorl site of psy538, and the reporter gene lacz with promoter pll was cloned into smai site of recombinant piasmid. both vp3 gene and lacz gene were cloned into noti site of psy681, recombinan fpv transfer vector containing vp3 gene of gpv was obtained. the result is basis of construction of recombinant fpv expressing vp3 gene of gpv and gpv genetic engineering vaccine
本研究另從含有gpvh1分離株主要結構蛋白vp3基因的重組質粒pproexhtb - vp3中切取gpvh1株vp3基因片段,將其亞克隆于psy538的ecori位點,然後將帶有痘苗病毒啟動子p11的lacz報告基因平端克隆于上述重組子的smai位點,再用noti切下同時含有vp3基因和lacz報告基因的片段,再亞克隆于psy681的noti位點,構建出含有vp3基因的重組禽痘病毒轉移載體,為構建表達vp3基因的重組禽痘病毒從而制備gpv基因工程疫苗奠定了基礎。Secondly, the hyaluronate lyase gene ( hyl ) was cloned into the vector puc19 by pcr using the total ona sample ofs. equi as template and partially sequenced, too
Equi的總dna為模板,通過pcr方法,克隆了透明質酸分解酶基因( hyl ) ,測序后連接到表達載體pse380的trc啟動子下游,構建表達質粒pse380 - hyl 。In this study, we designed a pair of primers based on the sequence of the upstream and downstream of chicken il - 2 gene. about 600 bp chicken il - 2 cdna fragment was cloned from cona - stimulated chicken splenocytes by reverse transcription - polymerase chain reaction ( rt - pcr ) and was subcloned into puc18 vector. recombinant clone was demonstrated by restriction enzyme digestion and dna sequencing. next, we construct recombinant plasmid pproex ? t - il - 2. the cdna of chicken il - 2 gene was subcloned into bamh i / hind iii sites of vector. the recombinant plasmid pproex ? t - il - 2 was transformed into e. coli dh5a and the bacteria was induced with iptg. it was demonstrated by sds - page and western blot that a 18kda protein which was equal to chicken il - 2 protein in molecular weight was expressed in e. coli dh5a. the expression level was up to 30 % of the total bacterial proteins. the purified protein was used to prepare the antibody against chicken il - 2 protein
經酶切鑒定及dna序列測定,該基因為雞il - 2基因,其序列與sundick等報道的完全一致。在此基礎上,我們把雞il - 2基因亞克隆到大腸桿菌原核表達載體pproex ~ ( tm ) ht中,構建重組表達質粒並進行確證性序列測定,重組質粒測序結果表明將編碼雞il - 2成熟蛋白的基因正確地插入到原核表達載體pproex ~ ( tm ) ht的目的位點。重組質粒轉化受體菌dh5後用iptg於37進行誘導培養, sds - page和westernblot分析顯示,表達的雞il - 2融合蛋白分子量約為18kda ,表達的融合蛋白經薄層掃描發現目的蛋白表達量約占菌體蛋白的30 。In this study, two kinds of specific promoter of b. thuringiensis : cry 3a promoter and btl - btll promoter were chosen to construct fusion genes to drive the expression of gfp gene in b. thuringiensis strain
本研究在構建蘇雲金芽胞桿菌質粒克隆載體pbmbi105的基礎上,克隆了菌株ybt一1520的質粒,獲得6 . 6kb的dna片段。The product of pcr named vp6 is approximate 1. 3kb in length. the vp6 gene was cloned into pmd18 - t vector and sequenced
將其插入克隆載體質粒pmd18 - t的ecorv酶切位點處,構建重組質粒pmd18 - t ? vp6 。Reconstruction of antisense pcdnas - doc - 1r plasmid after transformation the pmeiss - fl3 - doc - 1r plasmid into e. coli cells, we have extracted the plasmid and cut doc - 1r fragment using xho i to get a 975 bp doc - 1r cdna fragment
Pcdna3 - doc - 1r反義重組質粒的構建將pme18s - fl3 - doc - 1r質粒轉化后進行質粒提取。用xhoi限制性內切酶從克隆載體上切取975bp的doc - 1rcdna片段。分享友人