右轉因素 的英文怎麼說
中文拼音 [yòuzhuǎnyīnsù]
右轉因素
英文
right turn factor-
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。The hwtx - i gene was chemically synthesized according to its known cdna sequence, the gene was inserted into vector ppic9k which contained aoxj promotor and the sequence of a secreting signal peptide - a - factor, the cloning ppic9k / hwtx - i was constructed and confirmed by two - step pcr and dna sequence analysis, then it was transformed into host strain gs115, a his + muts cell line was screened and multicopy transformants were screened by various g418 concentrations, the multicopy transformant was named gh1. gh1 was cultivated in flasks. after 6 days of induction by 0. 5 % methanol, the supernatant was checked by 16. 5 % tricine - sds page, which showed there was a band in the position of 3. 5 - 6. 1kd, then it was isolated and desalted by ultrofiltration followed by ion exchange of cm column, after reverse phase hplc of ci8 and vacuum drying, the purified rhwtx - 1 was obtained which was proved to be correct recombinant hwtx - i by tricine sds - page, maldi - tof mass spectrometry, amino acid composition analysis, the n - terminal amino acid sequence and its biological activity, the final field of the purified rhwtx - i was about 80mg / l, accounting for 23. 6 % of it total secretory proteins
將帶有hwtx -基因的ppic9k經blgii線性化后，轉化酵母宿主菌gs115原生質體后經篩選陽性克隆並經表型鑒定為his ~ + mut ~ s酵母菌，進一步用遺傳毒素g418篩選多拷貝的轉化菌株，命名為gh1 ；將gh1甲醇酵母菌用0 . 5的甲醇誘導表達，發酵上清經90飽和度的（ nh _ 4 ） _ 2so _ 4沉澱， yw - 3 （ mwc03000 ）的超濾膜超濾，再經cm陽離子交換， c _ （ 18 ）反相hplc純化得到分子量為4kd左右的組分，其中4289 . 05的組分經質譜鑒定，氨基酸組成分析和序列測定為正確的表達產物，生物學活性表明其活性為天然毒素活性70 % ，表達量為80mg / l 。Right turn factor
右轉因素
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