sequencing gel 中文意思是什麼

sequencing gel 解釋
dna測序凝膠
  • sequencing : 測序，序列測定
  • gel : n. 凍膠，凝膠（體）；定型發膠。vi. （-ll-） 成凍膠，膠化。
  1. By means of discontinuous tris - hcl buffer system and poly aery 1 amide gel electrophoresis ( page ), zymogram of 23 species of 11 genera in 4 subfamilies mirinae, orthotylinae, phylinae and deraeocorinae are gained. by specific primer pcr and sequencing, 57 cyt b gene 433bp sequences are obtained from 34 species respectively belonging to 17 genera in 5 subfamilies mirinae, orthotylinae, phylinae, deraeocorinae, bryocorinae from 64 species of 23 genera which involved in this research, and 2 outgroup species of family anthocoridae as well. 1

    首次通過特異引物擴增和基因測序的研究方法從被試的5亞科23個屬的64種盲蝽中獲得了盲蝽亞科、合墊盲蝽亞科、葉盲蝽亞科、赤爪盲蝽亞科和蕨盲蝽亞科bryocorinae5個亞科17個屬的34種盲蝽以及外群花蝽科anthocoridae2種花蝽的cytb基因序列57條。
  2. 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。
  3. After electrophorised on 1 % agarose gel, the pcr production was purified with agarose gel dna extraction kit. the segment was ligated with vector pmd18 - t and then was tranformed into the competent cell of dh5 a. a construction mstnd - pmd18t was generated by inserting the sequence of 254bp into pmd18 - t vector and selecting the sense clones. positive clone was identified by three ways : endonuclease digestion, pcr and sequencing. the result showed that the cloned sequence coincides with the designed sequence. this construction was digested with nco i and xho i and ligated the pet28a ( + ) vector digested with the same enzymes using dna ligation kit. the production of ligation reaction was transformed into the competent cell of bl21 ( de3 ). after 12 - 16 hours of culture, several colnes appeared on the plate. some positive clones were selected to extract their plasmid. these plasmids were digested by nco i and xho i and indentified by pcr. a contraction, mstnd - pet28a was generated. the result showed that the cloned sequence coincides with the designed sequence

    F _ 1長38bp ， r _ 1長36bp ，其它片段均40bp長， f _ 1和r _ 1片段兩端分別加上限制性內切酶nco和xho的識別位點序列。用成對單鏈片段進行延伸反應，然後用其他單鏈片段作為引物，進行pcr擴增，用dna快速純化回收試劑盒回收所得254bppcr產物，與pmd18 - t載體連接、轉化dh _ 5 。受體菌感受態細胞，利用藍白斑遺傳學篩選法篩選陽性克隆，提取其質粒，採用nco和xho雙酶切鑒定，獲得了254bp的片段；用pmd18 - t載體上的特異引物rv - m和m13 - 47進行pcr鑒定，獲得300bp的片段。
  4. The nucleic acids of the all influenza viruses conducted in the research were extracted from the viruses propagated in specific - pathogen - free chicken embryos. all of the eight segments were amplified by rt - pcr, and the purified pcr products were done cycle sequencing with specific primers, furthermore, the sequencing products were purified and run gel on abi prism 377 dna sequencing machine. the specific primers of the eight genes for pcr and cycle sequencing were designed using the ohgo ( 4. 0 version ) and genedoc ( 2. 3 version ) software

    首先將實驗用毒株在spf雞胚中增殖，提取核酸，然後應用oligo （ 4 . 0版本）和gendoc （ 2 . 3版本）軟體設計h9n2aiv所有8個基因片段特異的pcr引物及序列測定引物，通過rt - pcr方法擴增所有毒株的各個基因片段，純化後用特異引物進行測序反應，反應產物純化后在abiprism377dna序列分析儀上進行序列測定，然後應用wisconsinsequenceanalysispackage （ gcg ， 10 . 2版本） 、 phylogeneticanalysisusingparsimony （ paup ， 4 . 0版本）和treeview （ 1 . 5版本）軟體進行序列的數據編輯、序列翻譯、進化樹繪制和遺傳演化分析。
  5. 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。
  6. In order to get small molecular g - protein rab3a, which serves to further investigate the interaction between rab3a and other proteins, we amplified the full coding region of rab3a cdna by polymerase chain reaction, using human placenta total cdna as template. the pcr products were recovered from gel electrophoresis and cloned into bamhi xhoi site of vector pyestrp2. the result of sequencing indicated that rab3a insertion fragment included its initiation and termination codons in 5 - and 3 - terminal, respectively

    為了獲取全長的小分子g -蛋白rab3a ，以用於研究rab3a與其他蛋白相互作用關系，本實驗以人胎盤總cdna為模板， pcr擴增到人rab3a cdna全編碼區。產物回收后克隆于質粒pyestrp2的bamhi xhoi位點，測序結果表明，本實驗獲得的rab3a cdna包含了起始和終止密碼子。
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