wild type virus 中文意思是什麼
wild type virus
解釋
野毒-
Construction of recombinant fowlpox virus coexpressing aiv ha and ndv f. for the construction of transfer vector pfgs11haf, aiv ha gene of f strain in puc18ha and ndv f gene of f48e8 strain in puc19f were removed and inserted into pfgs11. recombinant fowlpox viruses ( rfpv ) coexpressing aiv ha gene and ndv f gene were constructed by using different promoters of ps and pe / l. recombinant rfpvs were derived by dosper liposome - mediated transfection with the two transfer vectors on chicken embryo fibroblast ( cef ) monolayer cultures which were infected by wild type fpv chinese vaccine strain 282e4 3 - 4 hours earlier
Puc18ha和sk質粒同時經hind 、 kpn酶切后連接得中間質粒skha ;將質粒skha用bamhi酶切回收ha基因插入到插入載體pfgs11中的bamhi位點,通過酶切鑒定獲得了pfgs11ha ;將含ndvf基因的質粒puc19f用hind 、 sal酶切經klenow酶補平插入到經sma酶切后的skifn中pe / l啟動子下獲得中間質粒skf ,再將質粒skf和puc18質粒先分別用ecor 、 xho酶切klenow補平,后再共同用sac酶切連接得puc18pelf , sal酶切回收pe / l - f基因盒插入到pfgs11ha的sal位點,通過酶切鑒定獲得了pe / l - f與ps - ha同向的表達載體pfgs11haf 。 -
No trace of any newly expressed protein band was noticed in supernant as well as in the cells by sds - page, except the verification of the substitution of beta - galactosidase gene ( the lose of galactosidase protein band ), which is a selective marker of the wild - type virus. elisa test results suggested the expression of egf in cells, but not in culture supernant. the quantitative calculation suggested the expressed egf was about 6 - 7 u g ( as egf standard ) per flask ( 2 > < 106 cells ) in the cellular extract
將重組病毒rbmbacph - egf以10moi感染bmn細胞, 72小時后收集培養細胞和上清;培養上清和經超聲波處理的細胞樣品elisa檢測發現胞內樣品中存在能與egf抗體免疫反應的產物,粗略估計表達量約6 7 g 2 10 ~ 6個細胞(相當于egf標準品) ;重組病毒rbmbacph - egf穿刺接種5齡家蠶幼蟲,每隔24h收集蠶血淋巴,經elisa檢測發現第4天表達量最高,根據egf標準曲線計算蠶血淋巴的表達量約32 g ml ; elisa定性實驗還發現正常蠶血也存在與egf抗體間交叉反應的物質。
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