competent cell 中文意思是什麼
competent cell
解釋
感受態細胞-
Then the linked products were transformed into the high competent cell of e. coli dh5a. based on - complementation of the detective - galactosidase, positive recombinant clone were screened from x - gal plate
從引物和基因序列的比對分析結果看， zhyf006序列與上下游引物的配對比例分別為s4 -
Bp23 celb genes, b. pwnilus endoglucanase and b. polymyxa beta - 1, 4 - endoglucanase " genes, respectively. it was recognized as a new gene encoding for endoglucanase of b. mega terium. the recombinant plasmid tvchi ( pmd18 ~ t inserted with chitinase encoding gene from aplls ) and e. coli - bacillus shuttle vector physooplk were digested by ecori and sail completely, and the chitinase gene was ligated with shuttle vector, and the recombinant vector was used to transform b. megaterium ap25 competent cell
平板拮抗實驗同野生菌株相比，轉化子對麥長蠕抱菌的抑制作用最明顯，抑制百分數最高可達33 . 3 % ，而apll3和ap25分別是23 . 1 %和25 . 6 % ，同時轉化子對小麥紋枯病菌、棉花立枯病菌、棉花枯萎病菌和小麥的全蝕病菌也具有較為明顯的抑制作用。 -
Benzyl chloride were used for extracting genomic dna of aspergillus. niger 14, about 1. 5kb specific fragment was obtained from genomic dna of aspergillus. niger 14 by pcr amplification with primers ( forward primer5 " ataggcatcatgggcgtctct3 " reverse - primer5 " cagctaagcaaaacactccgc 3 designed according to the known sequences of the phytase gene in the gene bank and pyrobest ? dna polymerase, after ligated with pmd18 - tvector, transformated into e. colidh5a competent cell successfully. 3. nucleotide sequence analysis of the cloned fragment revealed the presence of the whole phya gene in pcr product
用氯化芐法提取了aspergillus . niger14 ~ #基因組dna ，根據genebank中已知的黑麴黴植酸酶基因序列設計出一對特異性引物（上游引物： 5 ataggcatcatgggcgtctc3下游引物： 5 cagctaagcaaaacactccgc3 ） ，採用pyrobest ~ （ tm ） dnapolymerase （高保真dna聚合酶） ，通過pcr方法從aspergillus . niger14 ~ #基因組dna中擴增出了預期的1 . 5kb左右的特異性產物，將其與pmd18 - tvector連接后，轉化e . colidh5菌株的感受態細胞，經質粒抽提、酶切鑒定，確認該目的產物已得到成功克隆。 -
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。 -
Sequence analysis shows that they share 98. 75 % similarity at the dna, and 98. 67 % at the protein level. ns2 gene was cloned into the multiple cloning sites of prokaryotic expression vector pgex - 6p - l. the recombinant plasmid pgex - 6p - ns2 was constructed and transformed to the competent cell bl21 ( de3 ) plyss, positive bacterium strain was induced by iptg
將ns2基因插入到原核表達性質粒pgex - 6p - 1的ecor 、 bamh多克隆位點之間，將重組原核表達質粒pgex - 6p - ns2轉化到bl21 （ de3 ） plyss感受態細胞中，獲得了表達ns2基因的陽性亞克隆重組子，在含amp的lb液體培養基中培養，經iptg誘導表達，用sds - page分析表達產物。 -
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。
分享友人