pET28GST-LIC大肠表达质粒 货号：P1087
来源:
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作者:shhebio
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发布时间: 2017-12-04
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148 次浏览
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分享到:
信息
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别名
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pET28GSTLIC
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启动子
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T7
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复制子
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pBR322
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终止子
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T7 terminator
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质粒大小
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7994bp
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质粒标签
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N-GST,N-6×HIS，N-thrombin
site，SacB
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原核抗性
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Kan
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真核抗性
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SacB
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克隆菌株
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DH5α
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培养条件
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37℃
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表达宿主
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大肠杆菌BL21(DE3)
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培养条件
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37℃，有氧，LB
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诱导方式
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IPTG和乳糖及其类似物
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5'测序引物
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T7（TAATACGACTCACTATAGGG）
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3'测序引物
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T7-ter（TGCTAGTTATTGCTCAGCGG）
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质粒宿主
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大肠杆菌
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质粒用途
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蛋白表达
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片段类型
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基因CDS
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片段物种
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空载体
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原核抗性
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卡那霉素
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简介
pET28GST-LIC质粒是一个大肠杆菌表达载体，T7启动子驱动GST和融合表达一起表达，载体上有SacB致死基因，可通过5%蔗糖来筛选阳性克隆。
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.
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