重介質選篩 的英文怎麼說
中文拼音 [zhòngjièzhíxuǎnshāi]
重介質選篩
英文
heavy media screen-
The total rna was isolated from pokeweed ( phytolacca americana ) leaves using the method of guanidine isothiocyante and used as template to amplify the total length and deleted mutant pokeweed antiviral protein ( pap ) gene by rt - pcr and then the pap gene was cloned into pgem - t vector. the sequencing results showed that pap gene had 99. 9 % identity comparing with the pap gene nucleotide sequence reported by lin et al ( 1991 ). the iptg - inducible expression vector containing the pap gene was constructed and transferred into e. coli bl21 ( de3 ) - plyss
將缺失型pap基因克隆到植物表達載體pbi121中，通過液氮冷凍法將重組質粒轉入農桿菌lba4404細胞中，然後採用葉盤法，在該農桿菌的介導下將pap基因導入普通煙草中，經過卡那黴素抗性篩選，最後獲得了轉pap基因的工程煙草植株，摩擦接種煙草花葉病毒（ tmv ） ，與非轉基因煙草相比，能夠推遲癥狀表現達2月之久，說明pap基因能夠在其它植物體內產生有活性的高抗病毒的蛋白質。Finally, a tranfer vector named as pltk - ha was constructed based on pltk - uni with insertion of ha gene of h3 subtype srv. the pltk - ha and prv bartha - k61 genomic dna were co - transfected into 50 - 70 % confluent vero cells in 6 - cm dishes. based on the expression of lacz gene, recombinant prv were selected and purified by blue - colored virus plaque
利用脂質體介導的方法，將pltk - ha與prvbartha - k61基因組共轉染于亞單層vero細胞，依據報告基因lacz在細胞中的表達，篩選藍色蝕斑的重組病毒，經數次蝕斑純化后， pcr鑒定、 western - blot分析。In this experiment hcv structural gene was amplified by polymerase chain reaction ( pcr ), and was inserted into baculovirus expression vector pfastbacl to construct a recombinant transposing vector pfbl - cee. the plasmid pfb 1 - cee was transformed into dh1 obac competent e. coli cells. high molecular weight dna was prepared from the overnight cultures from the selected e. coli colonies, which was recombinant baculovirus shuttle vector containing hcv structural gene, named bac - cee
本實驗用pcr擴增hcv結構區基因，克隆到桿狀病毒表達載體pfastbacl中，構建成重組轉座載體pfb1 - cee ，轉化dh10bac大腸桿菌感受態細胞，篩選陽性菌落，抽提大分子質粒dna ，獲得含hcv結構區基因的重組桿狀病毒穿梭載體bac - cee ，脂質體介導轉染sf9昆蟲細胞，出現細胞病變后，收集含有重組桿狀病毒顆粒的培養上消，重新感染sf9細胞，收集sf9細胞，進行12 . 5 sds -聚丙烯酰胺凝膠電泳，可見表達的蛋白條帶。
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