常用pGEX载体图谱.docx

1、常用pGEX载体图谱Rosetta系列的表达菌株可以提供T7 RNA聚合酶，它能表达PET系列载体上的外源基因。pGEX系列载体上的外源基因不需要T7 RNA聚合酶，普通的大肠杆菌经IPTG诱导即可表达Tac启动子是一组由Lac和trp启动子人工构建的杂合启动子，受Lac阻遏蛋白的负调节，它的启动能力比Lac和trp都强。其中Tac 1是由Trp启动子的35区加上一个合成的46 bp DNA片段(包括Pribnow 盒)和Lac操纵基因构成，Tac 12是由Trp的启动子-35区和Lac启动子的-10区，加上Lac操纵子中的操纵基因部分和SD序列融合而成蛋白标签：A myc tag is a

2、polypeptideprotein tagderived from thec-mycgene product that can be added to a protein usingrecombinant DNAtechnology. It can be used foraffinity chromatography, then used to separate recombinant, overexpressed protein from wild type protein expressed by the host organism. It can also be used in the

3、 isolation of protein complexes with multiple subunits.A myc tag can be used in many different assays that require recognition by anantibody. If there is no antibody against the studied protein, adding a myc-tag allows one to follow the protein with an antibody against the Myc epitope. Examples are

4、cellular localization studies by immunofluorescence or detection byWestern blotting.The peptide sequence of the myc-tag is (in 1- and 3-letter codes, respectively): N-EQKLISEEDL-C, N-Glu-Gln-Lys-Leu-Ile-Ser-Glu-Glu-Asp-Leu-C, where N stands forAmino-terminusand C stands forCarboxy terminus. The tag

5、is approximately 1202Daltonsin atomic mass and has 10 amino acids.It can be fused to theC-terminusand theN-terminusof a protein. It is advisable not to fuse the tag directly behind the signal peptide of a secretory protein, since it can interfere with translocation into thesecretory pathway.A monocl

6、onal antibody against the mycepitope, named 9E10, is available from the non-commercialDevelopmental Studies Hybridoma BankpGEX4T1载体基本信息出品公司:GE别名:pGEX-4T-1, pGEX4T1, pGEX 4T 1质粒类型:大肠杆菌蛋白表达载体表达水平:高拷贝启动子:Tac克隆方法:多克隆位点，限制性内切酶载体大小:4969 bp5 测序引物及序列:pGEX5: GGGCTGGCAAGCCACGTTTGGTG3 测序引物及序列:pGEX3: CCGGGAGCTG

7、CATGTGTCAGAGG载体标签:N-GST载体抗性:Ampicillin 氨苄备注:复制子是pMB1产品目录号:27-4580-01稳定性:瞬时表达 Transient组成型:诱导表达病毒/非病毒:非病毒pGEX4T1载体质粒图谱和多克隆位点信息原核生物DNA复制起始点，是DNA链上独特的具有起始DNA复制功能的碱基序列。大肠杆菌的复制起点包括OriC和OriHThrombin 凝血酶pGEX4T1载体简介pGEX4T1载体序列LOCUS pGEX-4T-1 4969 bp DNA circular SYNDEFINITION pGEX-4T-1ACCESSION KEYWORDS SOU

8、RCE ORGANISM other sequences; artificial sequences; vectors.COMMENT This file is created by Vector NTI COMMENT VNTAUTHORNAME|FEATURES Location/Qualifiers source 1.4969 /organism=pGEX-4T-1 /mol_type=other DNA promoter 184.212 /label=tac_promoter misc_feature 224.246 /label=M13_pUC_rev_primer gene 258

9、.977 /label=GST (variant) /gene=GST (variant) CDS 258.977 /label=ORF frame 3 misc_feature complement(1019.1041) /label=pGEX_3_primer promoter 1307.1335 /label=AmpR_promoter gene 1377.2237 /label=Ampicillin /gene=Ampicillin CDS 1377.2237 /label=ORF frame 3 rep_origin 2392.3011 /label=pBR322_origin mi

10、sc_feature 3309.4400 /label=lacI CDS 3441.4400 /label=ORF frame 3 promoter 4449.4478 /label=lac_promoter misc_feature 4492.4514 /label=M13_pUC_rev_primer promoter 4513.4531 /label=M13_reverse_primer CDS 4523.81 /label=ORF frame 2 misc_feature 4540.4695 /label=lacZ_a promoter complement(4543.4559) /l

11、abel=M13_forward20_primer misc_feature complement(4552.4574) /label=M13_pUC_fwd_primerpGEX4T2载体基本信息出品公司:GE别名:pGEX-4T-2, pGEX4T2, pGEX 4T 2质粒类型:大肠杆菌蛋白表达载体表达水平:高拷贝启动子:Tac克隆方法:多克隆位点，限制性内切酶载体大小:4970 bp5 测序引物及序列:pGEX5: GGGCTGGCAAGCCACGTTTGGTG3 测序引物及序列:pGEX3: CCGGGAGCTGCATGTGTCAGAGG载体标签:N-GST载体抗性:Ampicill

12、in 氨苄备注:复制子是pMB1产品目录号:27-4581-01稳定性:瞬时表达 Transient组成型:诱导表达病毒/非病毒:非病毒pGEX4T2载体质粒图谱和多克隆位点信息pGEX4T2载体简介pGEX4T2载体序列LOCUS pGEX-4T-2 4970 bp DNA circular SYNDEFINITION pGEX-4T-2ACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors.COMMENT This file is created by Vector NTI COMM

13、ENT VNTAUTHORNAME|FEATURES Location/Qualifiers source 1.4970 /organism=pGEX-4T-2 /mol_type=other DNA promoter 184.212 /label=tac_promoter misc_feature 224.246 /label=M13_pUC_rev_primer gene 258.978 /label=GST (variant) /gene=GST (variant) CDS 258.974 /label=ORF frame 3 misc_feature complement(1020.1

14、042) /label=pGEX_3_primer promoter 1308.1336 /label=AmpR_promoter gene 1378.2238 /label=Ampicillin /gene=Ampicillin CDS 1378.2238 /label=ORF frame 1 rep_origin 2393.3012 /label=pBR322_origin misc_feature 3310.4401 /label=lacI CDS 3442.4401 /label=ORF frame 1 promoter 4450.4479 /label=lac_promoter misc_feature 4493.4515 /label=M13_pUC_rev_primer promoter 4514.4532 /label=M13_reverse_primer CDS 4524.81 /label=ORF frame 3 misc_feature 4541.4696 /label=lacZ_a