isolated signal line 中文意思是什麼
isolated signal line
解釋
隔離信號線-
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 。
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