連結酶 的英文怎麼說
中文拼音 [liánjiē]
連結酶
英文
ligase-
The fusion protein was bactericidal active against staphylococcus aureus. in present study, we will truncate the none channel - forming do main, then attach the agrd to the pore - forming region ( k544 - i626 ) to construct a new engineered multidqmain protein machine - compact engineered peptide targeting staphylococcus aureus. such engineered peptide was constructed by linking the gene of staphylococcal agrd pheromone with the gene of c - terminal ( 1626 ) of colicin la pore - forming region ( k544 - i626 ) with site - directed mutation
利用點突變方法將金黃色葡萄球菌信息素agrd （ i型， ystcdfim ）的基因引入到大腸菌素fa梭基端1626基因上，並將限制性內切酶sacl酶切位點基因分別引入到大腸菌素fa的p4和k544上，通過酶切、膠回收、連接獲得含大腸菌素ia水性孔道結構域和金黃色葡萄球菌信息素agrd基因的重組質粒。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。Ligase an enzyme that catalyzes the bond formation between two substrates at the expense of the breakdown of atp or some other nucleotide triphosphate
連接酶：可將兩個底物結合在一起的酶，此過程需要atp或其他核苷三磷酸供能。There was no difference in other biologic characteristic of mscs between the two separation method, such as cell anchorage ratio and clone formation ratio. ( 2 ) plga film presented uniformity frame with no protuberance and fissure under scanning electron microscopy ( sem ). big aperture with smooth wall and average 400 m i n size running - through each other was observed in porous plga substrate, around the big aperture there were many round micropores about 5 m size. all of the structure were equal and uniform, which satisfied the further research work. ( 3 ) mscs adhesion at earlier time was promoted by biotiegenrafter 3h, cell number was ( 1. 5 0. 18 ) 105 in the plga film coated with biotiegen group, which was significantly higher than that in plga film group ( p < 0. 01 ) and higher than that in coverslip group ( p < 0. 05 ), which cell number was ( 1. 04 0. 21 ) 105. after 6h and 12h biotiegen could not promote cell adhesion, and cell proliferation and alkaline phosphatase ( alp ) activity were not promoted dramatically during 9 days. ( 4 ) cell adhesion was promoted by fibronectin or collagen type i
G ） i型膠原、纖維粘連蛋白促進細胞增殖，細胞接種后3 、 6 、 gd三個檢測時間點，實驗組細胞均明顯高於對照組。與1型膠原相比，纖維粘連蛋白刺激作用更強。 ） i型膠原、纖維粘連蛋白尚能誘導mscs細胞向成骨細胞分化，不僅表達成骨細胞標志物ocn 、 alp 、 opnmrna ，而且堿性磷酸酶活性明顯增高，堿性磷酸酶及鈣結節7第四軍醫大學博士學位論文一染色均強陽性， i型膠原組mscs細胞堿性磷酸酶活性較fn組更高，有顯著性差異；同時，兔疫組化染色表明，經纖維粘連蛋白作用的mscs1型膠原表達陽性。Levels of fasting blood glucose and 24h urinary microcontent of albumin 24 h malb were determined dynamically ; the serum glycosyl hemoglobin hba1c was determined after the last medication ; the ultrastructural changes of kidney were observed by transmission electron microscope ; the expressions of collagen, fibronctin, laminin ln, and the ecm metabolism influencing factors, including mmp2, tissue inhibitor of metalloproteinase timp2, transfer growth factor 1 tgf 1 in renal tissue were detected by immunohistological chemistry and image collecting analytical system
動態檢測各組大鼠空腹血糖fbg 24h尿微量白蛋白24h malb ，末次給藥后測定大鼠血漿糖化血紅蛋白hba1c透射電鏡觀察各組大鼠腎臟超微結構改變，應用免疫組化技術及圖像採集分析系統測定各組大鼠腎臟組織中型膠原c纖維連接蛋白fn層粘連蛋白ln的表達，測定影響ecm代謝的基質金屬蛋白酶2 mmp2基質金屬蛋白酶抑制劑2 timp2及轉化生長因子1 tgf 1的表達。The results of lauryl sodium sulfate - polyacrylamide gel electrophoreses ( sds - page ) of the aggregate precipitate and supernatant and the result of high - performance size - exclusion chromatography of the supernatant indicated that, by wrongly linked intermolecular disulfide bonds soluble bi - molecular and tri - molecular egg white lysozyme aggregate could be simultaneously formed except being renatured to native and active egg white lysozymes during the refolding procedure of denatured - reduced egg white lysozyme ; the aggregate precipitate could be further formed by the non - covalent bonds interaction between the soluble hi - molecular egg white lysozyme aggregates, and the soluble tri - molecular egg white lysozyme aggregate could still stay at the supernatant
沉澱和上清液的不連續十二烷基硫酸鈉聚丙烯酰胺凝膠電泳（ sds - page ）和高效凝膠排阻層析分析結果表明，還原脲變性蛋白溶菌酶在稀釋復性過程中除了能夠復性成天然態蛋白溶菌酶分子外，還會形成可溶的蛋白溶菌酶分子二聚體和三聚體，二聚體和三聚體主要是靠分子間二硫鍵的錯配連接而成的；可溶的蛋白溶菌酶分子二聚體之間通過非共價鍵相互作用而形成集聚體沉澱，而可溶的三聚體溶菌酶分子則仍處于復性液上清液中。The predicted vfcpkl protein with 493 amino acids contains all the structural characteristics of reported cdpks and there are four domains of a variable domain, a kinase domain, an autoinhibitory domain and a calmodulin - like domain from n - to c - terminal end
由編碼區推測的vfcpk1的493個氨基酸序列具有已報道的cdpk的所有典型結構特徵，由n端到c端可分為可變區、激酶區、連接區和調節區四個結構域。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。The results showed the soil enzymes activity and available nutrients in soybean continuous cropping could approach or higher than those of conventional fertilizeration after applying soybean specific fertilizer, which indicated soybean specific fertilizer could modulate the deteriorating soil enzymes and nutrients ecological surrondings, therefore it could overcome the soybean drop in production caused by continuous cropping
結果顯示施用大豆專用肥后，土壤酶活性和有效養分總體均能達到或高於輪作土壤常規施肥水平，這表明大豆專用肥能調控大豆連作所引起的土壤酶活性和養分惡化，最終使得大豆專用肥能克服大豆連作所引起的大幅度減產。The interesting gene fragment with ecori and noti were amplified by overlapping pcr, which inserted into vector plasmid ppic9k after degisted by ecori and noti, and the recombinant plasmid was transformed into competent dh5cc. positive clones were screened by pcr from the lb plate with amp. digesting analysis resulte shows that the interesting gene were inserted into the vector ppic9k with correct direction
目的基因經雙酶切后連接載體ppic9k ，然後導入大腸桿菌dh5中，在含氨卞青霉素（ amp ）的lb板上用pcr反應篩選出陽性菌落，雙酶切結果表明目的基因已插入載體中，且方向正確，測序結果進一步證明人巨細胞病毒重組基因表達質粒成功地克隆了目的基因片段。1. the heredity of strain yz - ii6 is quite stable because there is no any change for hydantoniase activity, even after passing the generations for 8 times
結果表明，該菌株的遺傳穩定性好，連續傳八代后海因酶活性仍無明顯變化。In this study, a new approach of plant virus - resistance was expored by making up the cdna of ppiv. nib gene was amplified from plasmid pysr, which is plant expression vector containing potyvirus y nib gene. the nib gene was fused with the fragment encoding the mature part of papaya proteinase iv
將番木瓜蛋白酶的cdna序列以全長、去除信號肽部分和成熟酶部分分別連接到細菌表達載體中，表達結果表明，只有含去除信號肽部分cdna的載體才能檢測到相應的表達的蛋白條帶。To construct eukaryotic expression vector of mbl gene with codon 54 point mutation, the target sequence in pgem - mbld plasmid, which conains mbl cdna with codon 54 mutant allele, was amplified by pcr. after the cdna fragement and plasmids pci - neo were prepared by digestion with sma i and sal i, the fragment was inserted into sma i and sal i site in pci - neo eukaryotic expression vector, and the recombinant vector, named pci - mbl54, was obtained. the pci - mbl54, digested with restriction enzymes, was found to contain the point mutation mbl cdna by agarose gel electrophoresis analysis
本實驗以ggc54gacmbl突變為研究對象，選用真核表達質粒pci - neo ，根據已構建好的含54位密碼子突變型mbl基因t載體的結構，設計合成新的引物， pcr擴增54位密碼突變型mbl基因，凝膠回收，雙酶切pcr產物和pci - neo質粒， t4連接酶連接，將前者克隆至後者的sma和sal位點，轉化大腸桿菌xl - 1blue ，氨芐選擇培養。Conclusion : by restriction enzyme secting, ligating, transforming, restriction enzyme analysis, and final dna sequencing, the pbd - i and pbd - ii gene were proved to be recombinated with the expression vector and the recombinated vector ppd - 1 and ppd2 were transformed successfully
結論：經過酶切、連結，構建成重組質粒ppd上、 ppd上，再經轉化、抽提質粒及酶切分析，最後經dna測序證實， rticr擴增的pbd i 、 pbd 11基因與piflpdt 」 xsi表達載體構建成功。After the pbd i and pbd ii gene ligated with the expression vector pinpoint ? a - 3, the recombinated plasmid ppd - 1 and ppd - 2 was transformed into jm109 strains, 4 positive clones were screened by restriction enzyme analysis, dna sequencing showed that two out of 4 positive clones inserted sequence of the constructed plasmid, which was the same as that of pbd - i and pbd - ii gene respectively, and its reading frame was correct, thus its could be used to express fusion protein
將pbd - 、 pbd -基因與表達載體pinpoint ~ （ tm ） xa - 3連結后獲得的重組質粒ppd - 1 、 ppd - 2轉化于大腸桿菌jm109中。經抽提質粒、酶切分析及pcr擴增，分別篩選到4個陽性克隆，將其中二個陽性克隆由測序分析，證實1個含pbd i基因片斷， 1個含pbd 11基因片斷，且閱讀框正確，可用於融合蛋白表達。So a lagre populations of relatively purified spermatids can be isolated from mouse testis by enzymatic digestion combined with discontinuous percoll gradient centrifugation method
因此採用組合酶消化結合非連續percoll梯度離。 g法，可有效地從小鼠睪丸中分離到較多較純的存活精子細胞。In this study, pichia pastoris system had been utilized for expression of fmdv 2c3abc gene which aimed for establishing a sensitive and specific molecular dignosis method. first, 2c and 3abc genes were amplified individually from p2 and 3abc postive clones and ligated together using pcr method, then this 2c3abc product was cloned into pgem - t easy vector and transformed e. coli dh5a competent cell. a postive recombinant plasmid which contained whole 2c3abc gene had been confirmed by pcr, enzyme digestion and sequencing. after that, the 2c3abc gene was sub - cloned into ppiczaa expression vector and transformed e. coli dh5 a competent cell and selected by zeocin ? antibiotic. the postive recombinant expression vector was linerized and electro - transformed pichia pastoris smd1168 competent cell. a recombinant pichia pastoris had been obtained by zeocin ? antibiotics selection and induced with 0. 5 % methanol for target protein expression. the expression product was analysised by sds - page and western blotting assay. the result sh owed that 2c3abc gene was expressed successfully in pichia pastoris and the product was a 95ku fusion protein which could be recognized by anti - fmdv serum. the amount of target protein was over 15 % of the total bacteria protein by gel thin layer scanning analysis. this research had supplied materials for establishing a fmd diagnosis method to differentiate infected animals from vaccinated animals
首先，用p2和3abc陽性克隆通過連接pcr方法獲得目的基因並將其克隆到pgem - teasy載體上，並轉化e colidh5a感受態細胞中，經pcr 、酶切以及測序證明得到了完整的2c3abc基因，並與國內外參考序列進行比較分析。然後，將目的基因亞克隆于ppiczaa表達載體並轉化大腸桿菌dh5a ，以zeocin ~ （ tm ）抗性篩選陽性克隆，大量提取重組表達質粒並用pme酶線性化后電轉化入畢赤酵母smd1168感受態細胞，通過zeocin ~ （ tm ）抗生素梯度濃度篩選，獲得重組酵母用0 . 5甲醇誘導表達，通過sds - page電泳、 westernblotting分析，結果表明， 2c3abc基因在畢赤酵母中成功表達，其表達產物為一95ku的融合蛋白，並能被口蹄疫病毒陽性血清識別。Sequences flanking tn5 - 1063a can be recovered from the genome of mutant by excision, self - ligation and transfer to e. coli. the total dna of mutant was excised with ecori, which cut the genome frequently but not cut the transposon. after sequencing the self - ligated transpon, dna fragment flanking tn5 was obtained. the result showed 042bm - x1 contains a tn5 insertion in the gene smc00190, which function was unknown and was demonstrated to be related to salt tolerance by this study, and the gene was named as rst - 0x1
通過ecori酶切突變株基因組，得到完整的tn5 （含有在大腸桿菌中起始復制的oriv ）及其側翼的序列片段，該片段自連后轉化大腸桿菌，以tn5兩端已知的序列設計引物進行測序。 blast的分析測序結果表明， 042bm - x1和042bm - x2中tn5分別定位在苜蓿中華根瘤菌1021染色體上smc02682和smc00419基因內部，本實驗證明它們和042bm耐鹽相關，命名為rst - 0x1和rst - 0x2基因。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 。In this research, the gpv hl isolate was propagated with 13 day ' s duck embryos. a pair of primers gflgr used to arnplify vp3 gene was designed using oligo4. i software according to the whole nucleotide sequence of gpv b isolate published by zadori. the major structural protein vp3 gene was amplified from the dna of gpv hl isoiate by polymerase chain reaction ( pcr ), and then cloned into pmdl8 - t vecter
根據zadori等發表的gpvb株全基因核苷酸序列，藉助oligo4 . 1軟體設計了1對用以擴增主要結構蛋白vp3基因的引物gf / gr ，通過pcr技術，從病毒基因組dna中擴增出病毒主要結構蛋白vp3完整基因片段，經酶切鑒定后直接與pmd18 - t質粒載體連接。分享友人