bgl 中文意思是什麼

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。

Bgl bulgaria leva

Bgl保加利亞利瓦

The fragments were ligated directly to the pichia pastrois expression vector ppic9 to got ppic9 - e3and ppic9 - e8. vectors were amplificated in the e. coli dh5 a and were linearized with bgl ii. the linearized vctors were transformed into host strain gs115. the recombinated strain was selected though phynotype and pcrthe positive strain was induced with methyl alcohol and was selected by dot - elisa. the recombinated protein was detected with sds - page and western - blot as before

重組菌用甲醇誘導表達，用dot - elisa的方法篩選到表達量較高的菌株。將篩選出的菌株大量的誘導表達，對表達上清處理后，用sds - page和western - blot進行鑒定。同時，用hiprep16 10heparinff肝素親和柱對表達蛋白進行了初步的純化。

Though the expression level of bl21 ( de3 ) / pt221 - hyuh was lower than that of m15 / pqe60 - hyuh, the target protein of bl21 ( de3 ) / pt221 - hyuh was principally in soluble form while the objective protein of m15 / pqe60 - hyuh was principally in insoluble form. both of the products in the two strains showed biological activity, but the former is 2 times higher than the latter. the hyuc dna fragment was inserted into ppic3. 5k plasmid to construct the ppic3. 5k - hyuc recombinant plasmid which was then transduced into pichia pastoris gs115 cells after being linearized by bgl ii digestion

結果表明， sds - page分析在50kd處有一較強的表達帶，融合有分子伴侶的重組菌株bl21 （ de3 ） pt221 - hyuh與非融合表達的重組菌株m15 pqe60 - hyuh相比，乙內酰脲水解酶的表達量低一些，但其表達蛋白主要以可溶性形式存在，而m15 pqe60 - hyuh中表達蛋白則主要以包含體形式存在，且前者的酶活性是後者的2倍多。