multicopy 中文意思是什麼
multicopy
解釋
多拷貝-
Considering alcaligenes faecalis pencillin g acylase ( afpga ), which possesses the attractive characteristics for beta - lactam antibiotics conversions, the gene of pga was cloned into two expressing vector pkkfpga and psmlfpga. both the constructed plasmids psmlfpga and pkkfpga contained the pga gene, trc promoter and rrnb transcript terminator, differ in the replicon and antibotic marker, pkkfpga contained multicopy replicon ( cole 1 ) and ampicillin marker. while psmlfpga contained medium - copy replicon ( p15a ) and tetracycline marker
本文將糞產堿桿菌青霉素g酰化酶（ afpga ）基因構建表達載體pkkfpga和psmlfpga ， pkkfpga和psmlfpga均含trc啟動子、青霉素g酰化酶基因、 rrnb轉錄終止子，其中pkkfpga含有氨卞青霉素抗性基因和cole1高拷貝復制子；而psmlfpga含有四環素抗性基因和p15a中拷貝復制子。 -
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 。 -
Multicopy integrants were screened with g418 from pichia pastoris which contains recombinant plasmid, and induced with methanol to secrete interesting peptide. the supernate of pichia pastoris culture was analysed by sds - page and western blotting. a reactive band, which the apparent molecular weight is 36kd, can be detected with sheep anti - hcmv polyclonal antibodies
重組質粒轉化巴氏畢赤酵母， g418篩選出多拷貝插入的單克隆，甲醇誘導多拷貝插入的單克隆酵母細胞分泌目的蛋白，培養液上清經sds - page電泳分析，在蛋白質印跡中檢測到培養液上清有一表觀分子量為36kd ，能與羊抗hcmv多克隆抗體發生發應的條帶。 -
Multicopy clones were screened from vary levels of g418 - ypd plate and cultured and induced with methanol to secrete interesting peptide, which was identified by sds - page and western blotting
培養多拷貝插入的酵母細胞，用甲醇誘導目的蛋白的分泌， sds - page和westernblotting鑒定表達蛋白。
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