目胞赤 的英文怎麼說
中文拼音 [mùbāochì]
目胞赤
英文
infantile palpebral swelling-
First, to construct a recombinant plasmid pegfp - c - fos with c - fos promoter and egfp, and then transfect it into human bladder transitional cell carcinoma biu - 87 cell ; second, based on the changes of the expression of gfp in the biu - 87 cell which induced by the aconitine and hab toxins, the concentration of the hab toxins could be detected
目的:構建一個含c - fos啟動子和egfp報告基因的pegfp - c - fos重組質粒載體。體外轉染膀胱癌biu - 87細胞后,利用赤潮毒素作用后細胞表達綠色熒光蛋白的變化來檢測赤潮毒素,初步建立一種以細胞為基礎受體水平的赤潮毒素檢測方法。Methods : an artificial gene for fl extracellular domain cdna was synthesized by using favored genetic codons of pichia pastroris. by inserting human fl extracellular domain cdna coding 156 amino acid resi dues into pichia pastoris expression vector ppic9k containing aox1 promoter and the sequences of alpha secreting signal peptides, a recombinant expression plasmid ppic9k - fl was constructed, and integrated into the alcohol oxidase region of the host genome
為了提高外源基因的表達量,我們根據畢赤氏酵母偏愛密碼子人工合成了編碼fl胞外區156個氨基酸的cdna序列,目的序列被定向克隆到酵母分泌型表達載體ppic9k質粒上,構建ppic9k - fl表達質粒。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。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。Multi - copies insertion transformants were screened on g418 plates. the recombinant protein was proved to have biological activity of hydrolyzing n - carbamoylphenylalanine into phenylalanine through enzyme activity assay. the n - carbamoylase activity of recombinant was 2. 26 and 2. 15 times higher than that of arthrobacter bt801 and dh5a / puc18 - 169
將hyucdna片段連接到真核表達載體ppic3 . 5k上,經bgl酶切線性化,通過peg法轉化導入畢赤酵母gs115感受態細胞,利用g418抗性篩選得到12個插入多拷貝目的基因的轉化子。分享友人