菌落轉化 的英文怎麼說
中文拼音 [jūnlàzhuǎnhuà]
菌落轉化
英文
colony transformation-
Construction and expression of yeast engineering yaccine : s14 / gnsag " as transformed to yeast host straln x33 by means of electroporation after ppiczaa s, . aresag " as l ined by saci enzyme. the single fungus, as choose and dibble inocu1ating in we and am plate, the positive fungus was gro ' ing in rm but not in w, and was 6 inoculated in ypd which included zeocine 500ug / ml and 1000ug / ml. 5 transformers were ampl if ied by pcr, three is same with positive control
選取單個菌落分別點種到刪平板和md平板，找出在回d上生長正常， w上生長緩慢或不生長的菌落，即陽性菌落，再以陽性菌落分別塗布zeocine含量500ug加， 1000ug砌l的ypd平板，以高濃度的抗生素篩選高拷貝的酵母工程菌，在含500ug ml高濃度抗生素平板上獲得了15個轉化子，取其中5個進行pcr擴增，有3個擴增產物與陽性對照相同，說明此酵母細胞中已含有s hbsag融合片段，其中之一命名為pIn 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。3. the effect of sporulation - independent promotor on toxicity of natural strain in order to study the effect of sporulation - independent promotor ( p3a ), p3a was spliced with the cry1c gene, then inserted into the shuttle vector pht304, and then recombinated plasmid pbmb827 was obtained. after transferring pbmb827 into strain ybt - 1520, it was surprising that the transformants had almost no potency against all lepidopteran larvae tested
3非依賴芽胞形成icp的cry3a啟動子（ p _ （ 3a ） ）對野生菌株特性的影響帶p _ （ 3a ）和cry1c基因的重組質粒pbmb827轉入ybt - 1520 ，轉化子對所測昆蟲的毒力下降非常明顯，芽胞和晶體也很難脫落。Culture of mg7 hybridoma cells and detection of antigen - binding affinity of mg7 mab by elisa 2. construction and identification of mg7 recombinant phage antibody library mrna was isolated from cultured mg7 hybridoma cells and converted into cdna ; the variable fragments of heavy and light chain were separately amplified and assembled into scfvs with a specially constructed dna linker by pcr. the scfvs dma was ligated into the phagmid vector pcantabse and the ligated sample was transfered into competent e. co / / tg1 to generate a bacterial form of mg7recombinant phage antibody library ; volume and recombinant ratio of the library were evaluated by means of bacterial colony counts and restriction analysis ( ecor i and hind iii )
Mg _ 7重組噬菌體抗體庫的構建及鑒定從培養的mg _ 7雜交瘤細胞中提取並分離mrna ，反轉錄成cdna ；利用pcr分別擴增mg _ 7單抗的重鏈及輕鏈可變區基因，並通過? dna連接子將二者連接起來形成mg _ 7單鏈抗體基因；將mg _ 7單鏈抗體基因插入pcantab5e ；將連接產物轉化感受態tg1大腸桿菌，制備細菌形式的mg _ 7重組噬菌體抗體庫；通過菌落計數和限制性酶切分析（ ecor和hind ）評估mg _ 7重組噬菌體抗體庫的容量和重組率。After linerization, the recombinant plasmid was transformed into pic hia pastoris by electroporation, which then were cultured in md plate free of histidine, from which the positive colones were propagated
重組質粒線性化后，用電擊法將重組質粒轉化入巴氏畢赤酵母，在缺組氨酸的md板上篩選陽性菌落，然後用不同濃度的g418 ? ypd板篩選多拷貝插入單菌落。When cotransforming pgbd - nifa and ad fusion genomic library into saccharomyces cerevisiae pj69 - 4a, 109 candidates interacting with nifa had been selected by testing for the expression of the his3, ade2 and lacz reporter genes
誘餌質粒和文庫質粒共轉化釀酒酵母（ saccharomycescerevisiae ） pj69 - 4a ，通過檢測報告基因his3 、 ade2及lacz的表達進行篩選，初篩得到109個陽性酵母菌落。A pair of primers were designed and synthesized based on the published ge gene sequence of prv - rice strain for amplifying ge gene of prv min - a, yielding a 1. 7kb band. the segment was linked to puc19 plasma dna by means of t4 dna ligase, transformed into e. coli jm109 permissive cells, and incubated on lb fray containg amp, x - gal and iptg. small amount of plasma was extracted by base cleavaging for enzyme digest analysis and pcr, resulting in recombinant plasma puge dna containing prv ge
用t _ 4dna連接酶使ge基因與經bamhi 、 kpni同樣雙酶切的puc19質粒dna連接；用連接產物轉化大腸桿菌jml09感受態細胞，置含amp 、 x - gal和iptg的lb平板上培養12 20小時；挑取白色菌落於選擇性培養基擴大培養，堿裂解法小量提取質粒dna ，並進行酶切分析鑒定，結果獲得整合有prvge基因的重組質粒pugedna ，並與其它prv分離株進行ge基因序列同源性分析。Selected one of the 14 strains - s93, s93 dna was digested partially with sau3a i and 2 ~ 3kb fragments were collected and inserted into puc 18, then transformed into dh5 a. filtering the clone with hybridization in situ, a 1 kb frament clone has been cloned
使用sau3ai對基因組dna進行不完全酶切，回收2 3kb片段，與puc18質粒連接轉化大腸桿菌，利用地高辛標記探針，使用菌落原位雜交篩選轉化子；篩選到包含有約1kb外源片段的轉化子。In y2h, rapgap from c. elegans was used as bait to screen c. elegans cdna library. after - lth screen, 63 colonies were checked
在酵母雙雜交系統中，將ppc97 rpg 」與ppc86七dna文庫共轉化入酵母菌y190細胞，經過一lth篩選后，有63個菌落擬為陽性菌落。Objective : to clone and sequence the cdna encoding metalloproteinase from the venom of agkistrodon acutus from northen mountain area of guangxi province. methods : one step method was used to extract total rna from the venom of agkistrodon acutus found in northern mountain area of guangxi province. different kinds of cdna encoding metalloproteinase were amplified by one step method ( rt - pcr and pcr reactions occurred in the same tube ) using different primers
方法：從桂北五步蛇毒腺中抽提總rna ，利用不同的引物，採用一步法（ rt - pcr和pcr在同一管內進行）擴增出不同的dna條帶，利用平端連接的方法將pcr擴增產物克隆至pgem - teasy載體，轉化大腸桿菌jm109 ，挑選白色菌落提取質粒，用pcr對其進行鑒定，直接利用純化pcr產物或提取陽性菌落質粒進行測序。In this experiment hcv structural gene was amplified by polymerase chain reaction ( pcr ), and was inserted into baculovirus expression vector pfastbacl to construct a recombinant transposing vector pfbl - cee. the plasmid pfb 1 - cee was transformed into dh1 obac competent e. coli cells. high molecular weight dna was prepared from the overnight cultures from the selected e. coli colonies, which was recombinant baculovirus shuttle vector containing hcv structural gene, named bac - cee
本實驗用pcr擴增hcv結構區基因，克隆到桿狀病毒表達載體pfastbacl中，構建成重組轉座載體pfb1 - cee ，轉化dh10bac大腸桿菌感受態細胞，篩選陽性菌落，抽提大分子質粒dna ，獲得含hcv結構區基因的重組桿狀病毒穿梭載體bac - cee ，脂質體介導轉染sf9昆蟲細胞，出現細胞病變后，收集含有重組桿狀病毒顆粒的培養上消，重新感染sf9細胞，收集sf9細胞，進行12 . 5 sds -聚丙烯酰胺凝膠電泳，可見表達的蛋白條帶。The ubi - sl - tocs fragment was taken out, inserted into the multi - cloning site of pcambia1300 vector, transformed into jm109 strain finally, positive colonies were screened on lb plate ( 60 g / ml kan added ). the result of pcr and enzyme digestion of plasmid proved that recombinatin vector was obtained ( named pcusaib4 and pcusaibu )
把擴增產物分別通過clai和bamhi酶切純化，連接到用clai和bamhi切去gfpml基因的中間表達載體pugfpocs中，轉化大腸桿菌jm109 ，在含amp （ 100 g / ml ）的lb抗性平板上篩選到了的陽性菌落。Main works and their important results are showed as following. firstly, the cdna encoding bullfrog gh is amplified by use of the total rna, extracted from adult bullfrog ' s pituitary, as the template, pi ( 5 ' - cggatcc atg gct tca ggg tta ggc ) and p2 ( 5 ' - cgaattc tta aaa ggt gca gtt gct ) as the primer pair and one particular cdna product, 660bp, was obtained after the rt - pcr - the product was purified by using dna agarose gel extraction method. after the purified cdna and pmd18 - t vector were ligated at 16 over night, the ligation products were transformed into e. coli strain dh5a and then the transformants were screened by clone pcr method
首先，以成體牛蛙的腦垂體總rna為模板，進行rt ? pcr擴增得到約660bp的特異性pcr產物帶，切下瓊脂糖凝膠中的特異產物帶進行cdna膠回收，將cdna與pmd18 ? t載體進行t ? a克隆連接並轉化到dh5 e . coli后，進行菌落pcr 、質粒酶切鑒定，篩選出陽性菌株，測序結果經blast分析，與已報道的牛蛙生長激素基因高度同源，證實此陽性克隆為牛蛙生長激素基因轉化子，命名為pbfgh 。These results are very important for us to further understand the genetic background, biological characteristics, evolutionary rule and the anti - schistosomajaponicum mechanism of microtus fortis at the molecular levels. the specific base changes of the dna fragme nt between the two subspecies are probably correlative with the animal immigration, survival conditions, and species evolution. the cdna library of microtus fortis bone marrow cells was transferred in situ to nylon membrane, which was divided into eight equals ( ga ~ - gh )
利用已經建立的東方田鼠骨髓細胞質粒cdna文庫，將cdna文庫轉化菌落印跡至尼龍膜，將膜均分成8份（ ga gh ） ，制備基因池，分別培養、提取基因池質粒dna ，通過lipofect - 2000脂質體轉染技術，將基因池質粒dna導入hek293細胞， 48h后收集轉染細胞上清液，即條件培養基。分享友人