■ 基本信息
別名: | pET52b |
啟動(dòng)子: | T7 |
復(fù)制子: | pBR322 |
終止子: | T7 terminator |
質(zhì)粒分類: | 大腸桿菌載體；PET系列表達(dá)質(zhì)粒 |
質(zhì)粒大小: | 5227bp |
質(zhì)粒標(biāo)簽: | N-Strep-Tag II，N-HRV 3C，C-Thrombin，C-10*His |
原核抗性: | Amp |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37℃，有氧，LB |
表達(dá)宿主: | BL21(DE3) |
誘導(dǎo)方式: | IPTG或乳糖及其類似物 |
5'測(cè)序引物: | T7:TAATACGACTCACTATAGGG |
3'測(cè)序引物: | T7-ter:TGCTAGTTATTGCTCAGCGG |
■ 質(zhì)粒屬性
質(zhì)粒宿主: | 大腸桿菌 |
質(zhì)粒用途: | 蛋白表達(dá) |
片段類型: | ORF
|
片段物種: | 空載體
|
原核抗性: | Amp |
真核抗性: |
|
熒光標(biāo)記: |
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■ 質(zhì)粒簡(jiǎn)介
The pET-52b(+) vector carries an N-terminal Strep?Tag? II coding sequence (1) followed by a recognition site for the human rhinovirus (HRV) 3C protease. This protease is highly specific for cleavage of the sequence LEVLFQ↓GP (2), and is active at low temperatures (3). The multiple cloning region is followed by an optional thrombin recognition site and a C-terminal His?Tag? coding sequence. The presence of two “gentle elution" tags at both the N-terminus and C-terminus is ideal for dual purification strategies designed to isolate full-length fusion proteins (4). Unique restriction sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below the circle map. The f1 origin is oriented so that infection with the helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand.
質(zhì)粒只保證關(guān)鍵序列正確，不保證表達(dá)效果。
■ 質(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列請(qǐng)下載：
ZK191pET-52b大腸表達(dá)質(zhì)粒.txt
