■ 基本信息
別名: | pET28GSTLIC |
啟動(dòng)子: | T7 |
復(fù)制子: | pBR322 |
終止子: | T7 terminator |
質(zhì)粒大小: | 7998bp |
質(zhì)粒標(biāo)簽: | N-GST,N-6×HIS，N-thrombin site，SacB |
原核抗性: | Kan |
真核抗性: | SacB |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37℃，有氧，LB |
表達(dá)宿主: | 大腸桿菌BL21(DE3) |
誘導(dǎo)方式: | IPTG和乳糖及其類似物 |
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: | Kan |
真核抗性: |
|
熒光標(biāo)記: |
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■ 質(zhì)粒簡(jiǎn)介
pET28GST-LIC質(zhì)粒是一個(gè)大腸桿菌表達(dá)載體，T7啟動(dòng)子驅(qū)動(dòng)GST和融合表達(dá)一起表達(dá)，載體上有SacB致死基因，可通過(guò)5%蔗糖來(lái)篩選陽(yáng)性克隆。
The pET28GST-LIC vector was derived from expression plasmid pET28a-LIC (SGC) by inserting the GST-tag from pET41a (Novagen) into the XbaI and NcoI sites. It is used for T7 promoter driven expression of recombinant proteins with the addition of a 242 amino acid N-terminal fusion tag containing the 217 amino acid GST-tag protein followed by a 6X His followed by a thrombin cleavage site. Two stop codons are included in the vector at the C-terminal cloning site.Insertion of DNA sequence into the cloning/expression region is preformed using BD-Biosciences Infusion enzyme mediated directional recombination between complementary 15 nucleotide DNA sequences at the ends of the insert (PCR product) and BseRI linearized vector. Insertion of target sequence involves replacement of a SacB gene stuffer sequence, which provides for negative selection of the original plasmid on 5% sucrose.
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：
ZK164pET28GST-LIC大腸表達(dá)質(zhì)粒.txt
