helper plasmid 中文意思是什麼
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4. engineering dhqase ( arod ) - deficient e. coli mutant with a second copy of the arob gene gene targeting technique was used to disrupt the arod gene in e. coli chromosome. the mutant 31bk was engineered, in which homologous recombination of the arobkanr gene cassette into the arod locus ( arod : : arobkanr ) of the e. coli strain atcc31884 genome utilized the helper plasmid pkd46 with red system. the host cell 31bk lacked catalytic activity of dhqase ( arod ) and had a second copy of the arob gene, so it improved carbon flow into the quinic acid biosynthesis direction
構建宿主菌基因精確定位突變株31bk （ arod : : arobkan ~ r ）為了改變代謝途徑脫氫奎尼酸（ dhq ）分支點上的代謝流量，使之充分流向目的產物奎尼酸合成方向，利用基因打靶技術構建了31884宿主菌arod基因精確定位插入突變體，使dhq脫水酶（ dhqase ）失活，阻斷了碳代謝流流向芳香氨基酸生成的方向，同時用同源重組的方法將arob基因定位整合入染色體上，解除了限速酶對碳代謝流通過共同途徑到達dhq的阻遏影響，並減輕代謝負擔。 -
Pgxn217 was transferred into bradyrhizobium japonicum strain gx201 by triparental mating using the helper plasmid prk2073. marker exchange was achieved by selection on yma containing sm, km, gm, spc et al four kinds of antibiotics. mutants were confirmed by southern blotting and hybridization with the wild - type region and the tn5 fragment respectively
隨后，利用突變質粒pgxn217誘變慢生型大豆根瘤菌菌株gx201 ，獲得了gx201的突變體菌株gx217 ；將pgxn201的3 . 4kbecori片段與plarf3連接獲得亞克隆pgxn201cl ，然後將pgxn201cl導入突變體菌株gx217 ，構建了gx217的功能互補菌株rgx217 。 -
The interest gene was inserted in the - tha l. tho 1 multiple cloning sites of donor plasimd pfastbachtb of baculovirus expression system. after analysis by restriction endonuclease and pcr, the recombinant donor plasmid gpl - fast was transforn1ed to the competent celi dhi0bac which contalns the bacmid and the helper plasmid, the recombinant bacmid gpl - bac was acquired which would express the vpl of gpv strain h l
同時將該目的基因插入到桿狀病毒表達系統的供體質粒pf _ （ ast ） b _ （ ac ） htb的xba 、 xho多克隆位點間，經酶切、 pcr鑒定后，將重組的供體質粒gp1 - fast轉化到含有桿狀病毒和輔助質粒的dh10b _ （ ac ）感受態細胞中，獲得了表達gpvh1株vp1的重組桿狀病毒gp1 - bac 。
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