酵素識別 的英文怎麼說
中文拼音 [jiàosùzhìbié]
酵素識別
英文
enzyme recognition- 酵 : 動詞（發酵） ferment; leaven
- 素 : Ⅰ形容詞1 （本色; 白色） white 2 （顏色單純） plain; simple; quiet 3 （本來的; 原有的） native Ⅱ名...
- 識 : 識Ⅰ動詞[書面語] （記） remember; commit to memory Ⅱ名詞1. [書面語] （記號） mark; sign 2. （姓氏） a surname
- 別 : 別動詞[方言] （改變） change （sb. 's opinion）
- 酵素 : biocatalyst
- 識別 : 1 （辯別; 辯認） discriminate; distinguish; discern; tell the difference; spot 2 [計算機] identif...
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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。
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