pBad/Myc-His A

pBadMyc-His A载体基本信息

pBadMyc-His A载体质粒图谱和多克隆位点信息

pBadMyc-His A载体简介

pBAD/His和PBAD/Myc-His载体质粒是衍生于pBR322载体。载体设计用来在大肠杆菌中进行可调节，剂量依赖性的表达和纯化重组目的蛋白。使用大肠杆菌araBAD启动子（pBAD）增强了大肠杆菌重组蛋白可溶性表达的水平。pBAD/His和pBAD/Myc His载体上的调节蛋白AraC能够调控pBad启动子。

pBAD/Myc-His A,B,C 载体简介

The pBAD/His and pBAD/Myc-His plasmids are pBR322-derived expression vectors designed for regulated, dose-dependent recombinant protein expression and purification in E. coli. Optimum levels of soluble, recombinant protein are possible using the araBAD promoter (PBAD) from E. coli. The regulatory protein, AraC, is provided on the pBAD/His and pBAD/Myc-His vectors allowing regulation of PBAD.

The pBAD/Myc-His Kit provides all of the necessary reagents to express your protein in a tightly regulated fashion. The pBAD/Myc-His vector expresses native proteins or fusion proteins with a C-terminal tag. The vector provides:

l   The araBAD promoter for tightly regulated expression

l   Translation initiation signals optimized for E. coliexpression

l   C-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin or detection with an Anti-His(C-term) Antibody

l   C-terminal c-myc epitope for detection and analysis with an Anti-myc Antibody. Three vectors are provided (A, B, and C). Each has the C-terminal tag in a different reading frame relative to the multiple cloning site to simplify in-frame cloning of your gene.

L-阿拉伯糖调控表达

In the presence of L-arabinose, expression from PBAD is turned on while the absence of L-arabinose produces very low levels of transcription from PBAD (Lee, 1980; Lee et al., 1987). Uninduced levels are repressed even further by growth in the presence of glucose. Glucose reduces the levels of 3′,5′-cyclic AMP, thus lowering expression of the catabolite-repressed PBAD promoter (Miyada et al., 1984). By varying the concentration of L-arabinose, protein expression levels can be optimized to ensure maximum expression of soluble protein. In addition, the tight regulation of PBAD by AraC is useful for expression of potentially toxic or essential genes (Carson et al., 1991; Dalbey and Wickner, 1985; Guzman et al., 1992; Kuhn and Wickner, 1985; Russell et al., 1989; San Millan et al., 1989). For more information on the mechanism of expression and repression of the ara regulon, refer to Schleif, 1992.

pBadMyc-His A载体序列

ORIGIN

    1 AAGAAACCAA TTGTCCATAT TGCATCAGAC ATTGCCGTCA CTGCGTCTTT TACTGGCTCT

   61 TCTCGCTAAC CAAACCGGTA ACCCCGCTTA TTAAAAGCAT TCTGTAACAA AGCGGGACCA

  121 AAGCCATGAC AAAAACGCGT AACAAAAGTG TCTATAATCA CGGCAGAAAA GTCCACATTG

  181 ATTATTTGCA CGGCGTCACA CTTTGCTATG CCATAGCATT TTTATCCATA AGATTAGCGG

  241 ATCCTACCTG ACGCTTTTTA TCGCAACTCT CTACTGTTTC TCCATACCCG TTTTTTGGGC

  301 TAACAGGAGG AATTAACCAT GGATCCGAGC TCGAGATCTG CAGCTGGTAC CATATGGGAA

  361 TTCGAAGCTT GGGCCCGAAC AAAAACTCAT CTCAGAAGAG GATCTGAATA GCGCCGTCGA

  421 CCATCATCAT CATCATCATT GAGTTTAAAC GGTCTCCAGC TTGGCTGTTT TGGCGGATGA

  481 GAGAAGATTT TCAGCCTGAT ACAGATTAAA TCAGAACGCA GAAGCGGTCT GATAAAACAG

  541 AATTTGCCTG GCGGCAGTAG CGCGGTGGTC CCACCTGACC CCATGCCGAA CTCAGAAGTG

  601 AAACGCCGTA GCGCCGATGG TAGTGTGGGG TCTCCCCATG CGAGAGTAGG GAACTGCCAG

  661 GCATCAAATA AAACGAAAGG CTCAGTCGAA AGACTGGGCC TTTCGTTTTA TCTGTTGTTT

  721 GTCGGTGAAC GCTCTCCTGA GTAGGACAAA TCCGCCGGGA GCGGATTTGA ACGTTGCGAA

  781 GCAACGGCCC GGAGGGTGGC GGGCAGGACG CCCGCCATAA ACTGCCAGGC ATCAAATTAA

  841 GCAGAAGGCC ATCCTGACGG ATGGCCTTTT TGCGTTTCTA CAAACTCTTT TGTTTATTTT

  901 TCTAAATACA TTCAAATATG TATCCGCTCA TGAGACAATA ACCCTGATAA ATGCTTCAAT

  961 AATATTGAAA AAGGAAGAGT ATGAGTATTC AACATTTCCG TGTCGCCCTT ATTCCCTTTT

 1021 TTGCGGCATT TTGCCTTCCT GTTTTTGCTC ACCCAGAAAC GCTGGTGAAA GTAAAAGATG

 1081 CTGAAGATCA GTTGGGTGCA CGAGTGGGTT ACATCGAACT GGATCTCAAC AGCGGTAAGA

 1141 TCCTTGAGAG TTTTCGCCCC GAAGAACGTT TTCCAATGAT GAGCACTTTT AAAGTTCTGC

 1201 TATGTGGCGC GGTATTATCC CGTGTTGACG CCGGGCAAGA GCAACTCGGT CGCCGCATAC

 1261 ACTATTCTCA GAATGACTTG GTTGAGTACT CACCAGTCAC AGAAAAGCAT CTTACGGATG

 1321 GCATGACAGT AAGAGAATTA TGCAGTGCTG CCATAACCAT GAGTGATAAC ACTGCGGCCA

 1381 ACTTACTTCT GACAACGATC GGAGGACCGA AGGAGCTAAC CGCTTTTTTG CACAACATGG

 1441 GGGATCATGT AACTCGCCTT GATCGTTGGG AACCGGAGCT GAATGAAGCC ATACCAAACG

 1501 ACGAGCGTGA CACCACGATG CCTGTAGCAA TGGCAACAAC GTTGCGCAAA CTATTAACTG

 1561 GCGAACTACT TACTCTAGCT TCCCGGCAAC AATTAATAGA CTGGATGGAG GCGGATAAAG

 1621 TTGCAGGACC ACTTCTGCGC TCGGCCCTTC CGGCTGGCTG GTTTATTGCT GATAAATCTG

 1681 GAGCCGGTGA GCGTGGGTCT CGCGGTATCA TTGCAGCACT GGGGCCAGAT GGTAAGCCCT

 1741 CCCGTATCGT AGTTATCTAC ACGACGGGGA GTCAGGCAAC TATGGATGAA CGAAATAGAC

 1801 AGATCGCTGA GATAGGTGCC TCACTGATTA AGCATTGGTA ACTGTCAGAC CAAGTTTACT

 1861 CATATATACT TTAGATTGAT TTAAAACTTC ATTTTTAATT TAAAAGGATC TAGGTGAAGA

 1921 TCCTTTTTGA TAATCTCATG ACCAAAATCC CTTAACGTGA GTTTTCGTTC CACTGAGCGT

 1981 CAGACCCCGT AGAAAAGATC AAAGGATCTT CTTGAGATCC TTTTTTTCTG CGCGTAATCT

 2041 GCTGCTTGCA AACAAAAAAA CCACCGCTAC CAGCGGTGGT TTGTTTGCCG GATCAAGAGC

 2101 TACCAACTCT TTTTCCGAAG GTAACTGGCT TCAGCAGAGC GCAGATACCA AATACTGTCC

 2161 TTCTAGTGTA GCCGTAGTTA GGCCACCACT TCAAGAACTC TGTAGCACCG CCTACATACC

 2221 TCGCTCTGCT AATCCTGTTA CCAGTGGCTG CTGCCAGTGG CGATAAGTCG TGTCTTACCG

 2281 GGTTGGACTC AAGACGATAG TTACCGGATA AGGCGCAGCG GTCGGGCTGA ACGGGGGGTT

 2341 CGTGCACACA GCCCAGCTTG GAGCGAACGA CCTACACCGA ACTGAGATAC CTACAGCGTG

 2401 AGCTATGAGA AAGCGCCACG CTTCCCGAAG GGAGAAAGGC GGACAGGTAT CCGGTAAGCG

 2461 GCAGGGTCGG AACAGGAGAG CGCACGAGGG AGCTTCCAGG GGGAAACGCC TGGTATCTTT

 2521 ATAGTCCTGT CGGGTTTCGC CACCTCTGAC TTGAGCGTCG ATTTTTGTGA TGCTCGTCAG

 2581 GGGGGCGGAG CCTATGGAAA AACGCCAGCA ACGCGGCCTT TTTACGGTTC CTGGCCTTTT

 2641 GCTGGCCTTT TGCTCACATG TTCTTTCCTG CGTTATCCCC TGATTCTGTG GATAACCGTA

 2701 TTACCGCCTT TGAGTGAGCT GATACCGCTC GCCGCAGCCG AACGACCGAG CGCAGCGAGT

 2761 CAGTGAGCGA GGAAGCGGAA GAGCGCCTGA TGCGGTATTT TCTCCTTACG CATCTGTGCG

 2821 GTATTTCACA CCGCATATGG TGCACTCTCA GTACAATCTG CTCTGATGCC GCATAGTTAA

 2881 GCCAGTATAC ACTCCGCTAT CGCTACGTGA CTGGGTCATG GCTGCGCCCC GACACCCGCC

 2941 AACACCCGCT GACGCGCCCT GACGGGCTTG TCTGCTCCCG GCATCCGCTT ACAGACAAGC

 3001 TGTGACCGTC TCCGGGAGCT GCATGTGTCA GAGGTTTTCA CCGTCATCAC CGAAACGCGC

 3061 GAGGCAGCAG ATCAATTCGC GCGCGAAGGC GAAGCGGCAT GCATAATGTG CCTGTCAAAT

 3121 GGACGAAGCA GGGATTCTGC AAACCCTATG CTACTCCGTC AAGCCGTCAA TTGTCTGATT

 3181 CGTTACCAAT TATGACAACT TGACGGCTAC ATCATTCACT TTTTCTTCAC AACCGGCACG

 3241 GAACTCGCTC GGGCTGGCCC CGGTGCATTT TTTAAATACC CGCGAGAAAT AGAGTTGATC

 3301 GTCAAAACCA ACATTGCGAC CGACGGTGGC GATAGGCATC CGGGTGGTGC TCAAAAGCAG

 3361 CTTCGCCTGG CTGATACGTT GGTCCTCGCG CCAGCTTAAG ACGCTAATCC CTAACTGCTG

 3421 GCGGAAAAGA TGTGACAGAC GCGACGGCGA CAAGCAAACA TGCTGTGCGA CGCTGGCGAT

 3481 ATCAAAATTG CTGTCTGCCA GGTGATCGCT GATGTACTGA CAAGCCTCGC GTACCCGATT

 3541 ATCCATCGGT GGATGGAGCG ACTCGTTAAT CGCTTCCATG CGCCGCAGTA ACAATTGCTC

 3601 AAGCAGATTT ATCGCCAGCA GCTCCGAATA GCGCCCTTCC CCTTGCCCGG CGTTAATGAT

 3661 TTGCCCAAAC AGGTCGCTGA AATGCGGCTG GTGCGCTTCA TCCGGGCGAA AGAACCCCGT

 3721 ATTGGCAAAT ATTGACGGCC AGTTAAGCCA TTCATGCCAG TAGGCGCGCG GACGAAAGTA

 3781 AACCCACTGG TGATACCATT CGCGAGCCTC CGGATGACGA CCGTAGTGAT GAATCTCTCC

 3841 TGGCGGGAAC AGCAAAATAT CACCCGGTCG GCAAACAAAT TCTCGTCCCT GATTTTTCAC

 3901 CACCCCCTGA CCGCGAATGG TGAGATTGAG AATATAACCT TTCATTCCCA GCGGTCGGTC

 3961 GATAAAAAAA TCGAGATAAC CGTTGGCCTC AATCGGCGTT AAACCCGCCA CCAGATGGGC

 4021 ATTAAACGAG TATCCCGGCA GCAGGGGATC ATTTTGCGCT TCAGCCATAC TTTTCATACT

 4081 CCCGCCATTC AGAG

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