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别名 | pFastBac Dual |
启动子 | P10+PH |
复制子 | pUC |
终止子 | SV40 poly(A) signal |
质粒分类 | 广宿主系列,昆虫细胞载体 |
质粒大小 | 5238bp |
原核抗性 | Amp |
真核抗性 | Gen |
克隆菌株 | DH5α |
培养条件 | 37℃ |
表达宿主 | 昆虫细胞 |
诱导方式 | 无须诱导,瞬时表达 |
5'测序引物 | pFastbac-F: TATTCCGGATTATTCATACC |
3'测序引物 | pFastBac-R: ACAAATGTGGTATGGCTGA |
质粒宿主 | 昆虫细胞 |
质粒用途 | 蛋白表达 |
原核抗性 | 氨苄青霉素 |
真核抗性 | 庆大霉素 |
pFastBac Dual昆虫细胞表达载体包含有两个启动子,能够同时表达两个外源基因,一个表达基因是polyhedrin (PH)启动子控制,另一个外源基因是由p10启动子控制。
pFastBac™Dual 载体的特点是在单个载体上有两个启动子,用于在使用 Bac-to-Bac® 杆状病毒表达系统(Bac-to-Bac® Baculovirus Expression
System) 时在昆虫细胞中同时表达两个蛋白质。该载体具有两个强启动子,多角体蛋白启动子和p10 启动子,因此可实现高水平表达。包含 CAT和β-葡萄糖醛酸酶 (gus) 的表达对照。
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.
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