■ 基本信息
別名: | pFastBac Dual |
啟動(dòng)子: | P10+PH |
復(fù)制子: | pUC |
終止子: | SV40 poly(A) signal |
質(zhì)粒分類(lèi): | 廣宿主系列，昆蟲(chóng)細(xì)胞載體 |
質(zhì)粒大小: | 5238bp |
原核抗性: | Amp |
真核抗性: | Gen |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37度 |
表達(dá)宿主: | 昆蟲(chóng)細(xì)胞 |
誘導(dǎo)方式: | 無(wú)須誘導(dǎo)，瞬時(shí)表達(dá) |
5'測(cè)序引物: | pFastbac-F:TATTCCGGATTATTCATACC |
3'測(cè)序引物: | pFastBac-R:ACAAATGTGGTATGGCTGA |
■ 質(zhì)粒屬性
質(zhì)粒宿主: | 昆蟲(chóng)細(xì)胞 |
質(zhì)粒用途: | 蛋白表達(dá) |
片段類(lèi)型: | ORF
|
片段物種: | 多框空載
|
原核抗性: | Amp,Gen |
真核抗性: |
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熒光標(biāo)記: |
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■ 質(zhì)粒簡(jiǎn)介
pFastBac Dual昆蟲(chóng)細(xì)胞表達(dá)載體包含有兩個(gè)啟動(dòng)子，能夠同時(shí)表達(dá)兩個(gè)外源基因，一個(gè)表達(dá)基因是polyhedrin (PH)啟動(dòng)子控制，另一個(gè)外源基因是由p10啟動(dòng)子控制。
pFastBacDual 載體的特點(diǎn)是在單個(gè)載體上有兩個(gè)啟動(dòng)子，用于在使用 Bac-to-Bac? 桿狀病毒表達(dá)系統(tǒng)(Bac-to-Bac? Baculovirus Expression System) 時(shí)在昆蟲(chóng)細(xì)胞中同時(shí)表達(dá)兩個(gè)蛋白質(zhì)。該載體具有兩個(gè)強(qiáng)啟動(dòng)子，多角體蛋白啟動(dòng)子和p10 啟動(dòng)子，因此可實(shí)現(xiàn)高水平表達(dá)。包含 CAT和β-葡萄糖醛酸酶 (gus) 的表達(dá)對(duì)照。
The Bac-to-Bac? Vector Kit contains a pFastBac? 1 vector, as well an expression control vector, intended for use as part of the Bac-to-Bac? Baculovirus Expression System , which enables the efficent production of recombinant baculovirus for expression testing in insect cells. The Bac-to-Bac? System relies on generation of recombinant baculovirus by site-specific transposition in E. colirather than homologous recombination in insect cells (Figure 1). This system features:
Time-saving expression bacmid. With Bac-to-Bac?, the expression cassette of the pFastBac? vector recombines with the parent bacmid in DH10Bac? E. coli Competent Cells (not included with this vector kit) to form an expression bacmid. The bacmid is then transfected into insect cells for production of recombinant baculovirus particles.
Easy colony screening. The parent bacmid in DH10Bac? E. colicontains a segment of the lacZα gene. The lacZα gene is disrupted upon transposition of the expression cassette into the bacmid allowing for blue/white selection of recombinants. This makes identification of recombinant colonies easy.
■ 質(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列請(qǐng)下載：
ZK1045pFastBacDual昆蟲(chóng)雙框質(zhì)粒.txt
質(zhì)粒只保證關(guān)鍵序列正確，不保證表達(dá)效果。
