| 出品公司: | Stratagene（Agilent） |
|---|---|
| 载体名称: | pBD-GAL4-Cam |
| 质粒类型: | 酵母双杂交载体；酵母噬粒载体 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 启动子: | ADH1 |
| 载体大小: | 6491 bp |
| 5' 测序引物及序列: | -- |
| 3' 测序引物及序列: | -- |
| 载体标签: | 无 |
| 载体抗性: | 氯霉素Chloramphenicol |
| 真核筛选标记: | TRP1 |
| 克隆菌株: | XL1-Blue MRF' |
| 表达菌株: | YRG-2 |
| 备注: | 含a beta-gal报告基因。配套载体为pAD-GAL4-2.1 |
| 产品目录号: | #838401-13 |
| 组成型/诱导型: | 组成型 |
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pBD-GAL4-Cam载体质粒基本信息
pBD-GAL4-Cam质粒图谱载体图谱和pBD-GAL4-Cam载体序列质粒序列多克隆位点信息
pBD-GAL4-Cam质粒载体简介
酵母双杂交系统
Protein–protein interactions occur in many biological processes including replication, transcription, secretion, signal transduction, and metabolism. A fundamental question in the study of any protein is to identify proteins that interact with a given protein in vivo. Intense research efforts are focused on the identification of these proteins.
The GAL4 two-hybrid phagemid vector system, a eukaryotic
system to detect protein–protein interactions in vivo, provides a method for the rapid identification of genes encoding proteins that interact with a given protein (i.e., a bait protein).1,2 The system is based on the ability to separate eukaryotic transcriptional activators into two separate domains, the DNAbinding domain (BD) and the transcriptional activation domain (AD).
In theGAL4 two-hybrid phagemid vector system, proteins that interact with the bait protein are identified by generating hybrids of the yeast GAL4 BD and the bait protein (X) and the GAL4 AD and a library of proteins (Y). Neither hybrid protein is capable of initiating specific transcription of reporter genes in yeast in the absence of a specific interaction with the other hybrid protein.
When the hybrid protein X is expressed in yeast, the GAL4 BD binds X to specific DNA sequences in the yeast chromosome defined by the GAL1 or GAL4 upstream activating sequences (UASGAL1 or UASGAL4, respectively), which regulate the expression of a reporter gene. Binding of X to the UAS is not sufficient to initiate transcription of the reporter gene. When Y is expressed in yeast, the AD interacts with other components of the transcription machinery required to initiate transcription of the reporter gene. However, Y alone is not localized to the reporter gene UAS and therefore does not activate transcription of the reporter gene. When a specific interaction between X and Y localizes both the GAL4 BD and GAL4 AD to the reporter gene UAS, transcriptional activation of the reporter gene occurs (Figure 2B). The reporter genes in the GAL4 twohybrid phagemid vector system are β-galactosidase (lacZ) and histidine (HIS3).
双杂交载体pAD-GAL4-2.1和pBD-GAL4-Cam
The pAD-GAL4-2.1 phagemid vector contains a multiple cloning site (MCS) with BamH I, Nhe I, EcoR I, Xho I, Sal I, Xba I, Pst I, and Bgl II restriction sites. The pBD-GAL4-Cam phagemid vector contains an MCS with EcoR I, Srf I, Sma I, Xho I, Sal I, Xba I, and Pst I restriction sites. The unique EcoR I and Xho I cloning sites in the pAD-GAL4-2.1 vector make this vector compatible with the Agilent cDNA Synthesis Kit for the preparation of unidirectional cDNA libraries.
The unique EcoR I and Sal I cloning sites are used for the preparation of cDNA libraries in the pBD-GAL4 Cam phagemid vector because the Xho I site in the MCS is not unique. The unique BamH I, Nhe I, and EcoR I sites at the 5′ end and the Xho I, Sal I, Xba I, and Bgl II sites at the 3′ end of the DNA insert facilitate the transfer of DNA encoding the target protein into commonly used protein expression/purification vectors. Genomic DNA digested with Mbo I, BamH I, or Sau3A I and partially filled-in can be inserted into a partially filled-in Xho I site in the pAD-GAL4-2.1 phagemid vector. The Xba I site in the pAD-GAL4-2.1 and pBD-GAL4-Cam phagemid vectors is not unique and contains the UAG amber suppressor in the same translational reading frame as the GAL4 domain. DNA should therefore be inserted such that the Xba I site is not between the GAL4 domain and the DNA insert.
The pAD-GAL4-2.1 and pBD-GAL4 Cam phagemid vectors contain the pUC origin for replication and an f1 origin for production of single-stranded DNA (ssDNA) in E. coli. Single-stranded DNA can be used for DNA sequencing or site-directed mutagenesis. The pAD-GAL4-2.1 and pBDGAL4-Cam phagemid vectors contain ampicillin-resistance gene [β-lactamase (bla)] and chloramphenicol acetyltransferase genes, respectively, for selection with ampicillin and chloramphenicol in E. coli. The pADGAL4-2.1 and pBD-GAL4 Cam phagemid vectors contain the 2μ origin for replication. For selection in yeast, the pAD-GAL4-2.1 phagemid vector contains the LEU2 gene and the pBD-GAL4 Cam phagemid vector contains the TRP1 gene. The hybrid protein is expressed by the alcohol dehydrogenase 1 (ADH1) promoter (P ADH1) and is terminated by the ADH1 terminator (T ADH1).
pBD-GAL4-Cam质粒序列载体序列
pBD-GAL4-Cam, 6491 bp version 045002
NOTE: The following sequence has been verified for accuracy
at the junctions. The remainder of the sequence has been
obtained from existing data.
1 GCTTGCATGC AACTTCTTTT CTTTTTTTTT CTTTTCTCTC TCCCCCGTTG
51 TTGTCTCACC ATATCCGCAA TGACAAAAAA ATGATGGAAG ACACTAAAGG
101 AAAAAATTAA CGACAAAGAC AGCACCAACA GATGTCGTTG TTCCAGAGCT
151 GATGAGGGGT ATCTCGAAGC ACACGAAACT TTTTCCTTCC TTCATTCACG
201 CACACTACTC TCTAATGAGC AACGGTATAC GGCCTTCCTT CCAGTTACTT
251 GAATTTGAAA TAAAAAAAAG TTTGCTGTCT TGCTATCAAG TATAAATAGA
301 CCTGCAATTA TTAATCTTTT GTTTCCTCGT CATTGTTCTC GTTCCCTTTC
351 TTCCTTGTTT CTTTTTCTGC ACAATATTTC AAGCTATACC AAGCATACAA
401 TCAACTCCAA GCTTGAAGCA AGCCTCCTGA AAGATGAAGC TACTGTCTTC
451 TATCGAACAA GCATGCGATA TTTGCCGACT TAAAAAGCTC AAGTGCTCCA
501 AAGAAAAACC GAAGTGCGCC AAGTGTCTGA AGAACAACTG GGAGTGTCGC
551 TACTCTCCCA AAACCAAAAG GTCTCCGCTG ACTAGGGCAC ATCTGACAGA
601 AGTGGAATCA AGGCTAGAAA GACTGGAACA GCTATTTCTA CTGATTTTTC
651 CTCGAGAAGA CCTTGACATG ATTTTGAAAA TGGATTCTTT ACAGGATATA
701 AAAGCATTGT TAACAGGATT ATTTGTACAA GATAATGTGA ATAAAGATGC
751 CGTCACAGAT AGATTGGCTT CAGTGGAGAC TGATATGCCT CTAACATTGA
801 GACAGCATAG AATAAGTGCG ACATCATCAT CGGAAGAGAG TAGTAACAAA
851 GGTCAAAGAC AGTTGACTGT ATCGCCGGAA TTCGCCCGGG CCTCGAGCCC
901 GGGTCGACTC TAGAGCCCTA TAGTGAGTCG TATTACTGCA GCCAAGCTAA
951 TTCCGGGCGA ATTTCTTATG ATTTATGATT TTTATTATTA AATAAGTTAT
1001 AAAAAAAATA AGTGTATACA AATTTTAAAG TGACTCTTAG GTTTTAAAAC
1051 GAAAATTCTT GTTCTTGAGT AACTCTTTCC TGTAGGTCAG GTTGCTTTCT
1101 CAGGTATAGC ATGAGGTCGC TCTTATTGAC CACACCTCTA CCGGCATGCC
1151 GGCAAGTGCA CAAACAATAC TTAAATAAAT ACTACTCAGT AATAACCTAT
1201 TTCTTAGCAT TTTTGACGAA ATTTGCTATT TTGTTAGAGT CTTTTACACC
1251 ATTTGTCTCC ACACCTCCGC TTACATCAAC ACCAATAACG CCATTTAATC
1301 TAAGCGCATC ACCAACATTT TCTGGCGTCA GTCCACCAGC TAACATAAAA
1351 TGTAAGCTTT CGGGGCTCTC TTGCCTTCCA ACCCAGTCAG AAATCGAGTT
1401 CCAATCCAAA AGTTCACCTG TCCCACCTGC TTCTGAATCA AACAAGGGAA
1451 TAAACGAATG AGGTTTCTGT GAAGCTGCAC TGAGTAGTAT GTTGCAGTCT
1501 TTTGGAAATA CGAGTCTTTT AATAACTGGC AAACCGAGGA ACTCTTGGTA
1551 TTCTTGCCAC GACTCATCTC CATGCAGTTG GACGATATCA ATGCCGTAAT
1601 CATTGACCAG AGCCAAAACA TCCTCCTTAG GTTGATTACG AAACACGCCA
1651 ACCAAGTATT TCGGAGTGCC TGAACTATTT TTATATGCTT TTACAAGACT
1701 TGAAATTTTC CTTGCAATAA CCGGGTCAAT TGTTCTCTTT CTATTGGGCA
1751 CACATATAAT ACCCAGCAAG TCAGCATCGG AATCTAGAGC ACATTCTGCG
1801 GCCTCTGTGC TCTGCAAGCC GCAAACTTTC ACCAATGGAC CAGAACTACC
1851 TGTGAAATTA ATAACAGACA TACTCCAAGC TGCCTTTGTG TGCTTAATCA
1901 CGTATACTCA CGTGCTCAAT AGTCACCAAT GCCCTCCCTC TTGGCCCTCT
1951 CCTTTTCTTT TTTCGACCGA ATTAATTCGT AATCATGGTC ATAGCTGTTT
2001 CCTGTGTGAA ATTGTTATCC GCTCACAATT CCACACAACA TACGAGCCGG
2051 AAGCATAAAG TGTAAAGCCT GGGGTGCCTA ATGAGTGAGG TAACTCACAT
2101 TAATTGCGTT GCGCTCACTG CCCGCTTTCC AGTCGGGAAA CCTGTCGTGC
2151 CAGCTGGTTC TTTCCGCCTC AGGACTCTTC CTTTTTCAAT ATTATTGAAG
2201 CATTTATCAG GGTTATTGTC TCATGAGCGG ATACATATTT GAATGTATTT
2251 AGAAAAATAA ACAAATAGGG GTTCCGCGCA CATTTCCCCG AAAAGTGCCA
2301 CCTGACGCGC CCTGTAGCGG CGCATTAAGC GCGGCGGGTG TGGTGGTTAC
2351 GCGCAGCGTG ACCGCTACAC TTGCCAGCGC CCTAGCGCCC GCTCCTTTCG
2401 CTTTCTTCCC TTCCTTTCTC GCCACGTTCG CCGGCTTTCC CCGTCAAGCT
2451 CTAAATCGGG GGCTCCCTTT AGGGTTCCGA TTTAGTGCTT TACGGCACCT
2501 CGACCCCAAA AAACTTGATT AGGGTGATGG TTCACGTAGT GGGCCATCGC
2551 CCTGATAGAC GGTTTTTCGC CCTTTGACGT TGGAGTCCAC GTTCTTTAAT
2601 AGTGGACTCT TGTTCCAAAC TGGAACAACA CTCAACCCTA TCTCGGTCTA
2651 TTCTTTTGAT TTATAAGGGA TTTTGCCGAT TTCGGCCTAT TGGTTAAAAA
2701 ATGAGCTGAT TTAACAAAAA TTTAACGCGA ATTTTAACAA AATATTAACG
2751 CTTACAATTT ACGCGCCTAA ATTATGCGGC CGCATATAAC TGGATTAATG
2801 AATCGGCCAA CGCGCGGGGA GAGGCGGTTT GCGTATTGGG CGCTCTTCCG
2851 CTTCCTCGCT CACTGACTCG CTGCGCTCGG TCGTTCGGCT GCGGCGAGCG
2901 GTATCAGCTC ACTCAAAGGC GGTAATACGG TTATCCACAG AATCAGGGGA
2951 TAACGCAGGA AAGAACATGT GAGCAAAAGG CCAGCAAAAG GCCAGGAACC
3001 GTAAAAAGGC CGCGTTGCTG GCGTTTTTCC ATAGGCTCCG CCCCCCTGAC
3051 GAGCATCACA AAAATCGACG CTCAAGTCAG AGGTGGCGAA ACCCGACAGG
3101 ACTATAAAGA TACCAGGCGT TTCCCCCTGG AAGCTCCCTC GTGCGCTCTC
3151 CTGTTCCGAC CCTGCCGCTT ACCGGATACC TGTCCGCCTT TCTCCCTTCG
3201 GGAAGCGTGG CGCTTTCTCA TAGCTCACGC TGTAGGTATC TCAGTTCGGT
3251 GTAGGTCGTT CGCTCCAAGC TGGGCTGTGT GCACGAACCC CCCGTTCAGC
3301 CCGACCGCTG CGCCTTATCC GGTAACTATC GTCTTGAGTC CAACCCGGTA
3351 AGACACGACT TATCGCCACT GGCAGCAGCC ACTGGTAACA GGATTAGCAG
3401 AGCGAGGTAT GTAGGCGGTG CTACAGAGTT CTTGAAGTGG TGGCCTAACT
3451 ACGGCTACAC TAGAAGGACA GTATTTGGTA TCTGCGCTCT GCTGAAGCCA
3501 GTTACCTTCG GAAAAAGAGT TGGTAGCTCT TGATCCGGCA AACAAACCAC
3551 CGCTGGTAGC GGTGGTTTTT TTGTTTGCAA GCAGCAGATT ACGCGCAGAA
3601 AAAAAGGATC TCAAGAAGAT CCTTTGATCT TTTCTACGGG GTCTGACGCT
3651 CAGTGGAACG AAAACTCACG TTAAGGGATT TTGGTCATGA GATTATCAAA
3701 AAGGATCTTC ACCTAGATCC TTTTAAATTA AAAATGAAGT TTTAAATCAA
3751 TCTAAAGTAT ATATGAGTAA ACTTGGTCTG ACAGTTACCA ATGCTTAATC
3801 AGTGAGGCAC CTATCTCAGC GATCTGTCTA TTTCGTTCAT CCATAGTTGC
3851 CTGACTCCCC GTCGTGTAGA TAACTACGAT ACGGGAGGGC TTACCATCTG
3901 GCCCCAGTGC TGCAATGATA CCGCGAGACC CACGCTCACC GGCTCCAGAT
3951 TTATCAGCAA TAAACCAGCC AGCCGCCTGT GACGGAAGAT CACTTCGCAG
4001 AATAAATAAA TCCTGGTGTC CCTGTTGATA CCGGGAAGCC CTGGGCCAAC
4051 TTTTGGCGAA AATGAGACGT TGATCGGCAC GTAAGAGGTT CCAACTTTCA
4101 CCATAATGAA ATAAGATCAC TACCGGGCGT ATTTTTTGAG TTGTCGAGAT
4151 TTTCAGGAGC TAAGGAAGCT AAAATGGAGA AAAAAATCAC TGGATATACC
4201 ACCGTTGATA TATCCCAATG GCATCGTAAA GAACATTTTG AGGCATTTCA
4251 GTCAGTTGCT CAATGTACCT ATAACCAGAC CGTTCAGCTG GATATTACGG
4301 CCTTTTTAAA GACCGTAAAG AAAAATAAGC ACAAGTTTTA TCCGGCCTTT
4351 ATTCACATTC TTGCCCGCCT GATGAATGCT CATCCGGAAT TACGTATGGC
4401 AATGAAAGAC GGTGAGCTGG TGATATGGGA TAGTGTTCAC CCTTGTTACA
4451 CCGTTTTCCA TGAGCAAACT GAAACGTTTT CATCGCTCTG GAGTGAATAC
4501 CACGACGATT TCCGGCAGTT TCTACACATA TATTCGCAAG ATGTGGCGTG
4551 TTACGGTGAA AACCTGGCCT ATTTCCCTAA AGGGTTTATT GAGAATATGT
4601 TTTTCGTCTC AGCCAATCCC TGGGTGAGTT TCACCAGTTT TGATTTAAAC
4651 GTGGCCAATA TGGACAACTT CTTCGCCCCG TTTTCACCAT GGGCAAATAT
4701 TATACGCAAG GCGACAAGGT GCTGATGCCG CTGGCGATTC AGGTTCATCA
4751 TGCCGTTTGT GATGGCTTCC ATGTCGGCAG AATGCTTAAT GAATTACAAC
4801 AGTACTGCGA TGAGTGGCAG GGCGGGGCGT AATTTTTTTA AGGCAGTTAT
4851 TGGTGCCCTT AAACGCCTGG TTGCTACGCC TGAATAAGTG ATAATAAGCG
4901 GATGAATGGC AGAAATTCGA AAGCAAATTC GACCCGGTCG TCGGTTCAGG
4951 GCAGGGTCGT TAAATAGCCG CTTATGTCTA TTGCTGGTTT ACCGGTTTAT
5001 TGACTACCGG AAGCAGTGTG ACCGTGTGCT TCTCAAATGC CTGAGGCCAG
5051 TTTGCTCAGG CTCTCCCCGT GGAGGCTAAG AAACCATTAT TATCATGACA
5101 TTAACCTATA AAAATAGGCG TATCACGAGG CCCTTTCGTC TCGCGCGTTT
5151 CGGTGATGAC GGTGAAAACC TCTGACACAT GCAGCTCCCG GAGACGGTCA
5201 CAGCTTGTCT GTAAGCGGAT GCCGGGAGCA GACAAGCCCG TCAGGGCGCG
5251 TCAGCGGGTG TTGGCGGGTG TCGGGGCTGG CTTAACTATG CGGCATCAGA
5301 GCAGATTGTA CTGAGAGTGC ACCATAACGC ATTTAAGCAT AAACACGCAC
5351 TATGCCGTTC TTCTCATGTA TATATATATA CAGGCAACAC GCAGATATAG
5401 GTGCGACGTG AACAGTGAGC TGTATGTGCG CAGCTCGCGT TGCATTTTCG
5451 GAAGCGCTCG TTTTCGGAAA CGCTTTGAAG TTCCTATTCC GAAGTTCCTA
5501 TTCTCTAGCT AGAAAGTATA GGAACTTCAG AGCGCTTTTG AAAACCAAAA
5551 GCGCTCTGAA GACGCACTTT CAAAAAACCA AAAACGCACC GGACTGTAAC
5601 GAGCTACTAA AATATTGCGA ATACCGCTTC CACAAACATT GCTCAAAAGT
5651 ATCTCTTTGC TATATATCTC TGTGCTATAT CCCTATATAA CCTACCCATC
5701 CACCTTTCGC TCCTTGAACT TGCATCTAAA CTCGACCTCT ACATTTTTTA
5751 TGTTTATCTC TAGTATTACT CTTTAGACAA AAAAATTGTA GTAAGAACTA
5801 TTCATAGAGT GAATCGAAAA CAATACGAAA ATGTAAACAT TTCCTATACG
5851 TAGTATATAG AGACAAAATA GAAGAAACCG TTCATAATTT TCTGACCAAT
5901 GAAGAATCAT CAACGCTATC ACTTTCTGTT CACAAAGTAT GCGCAATCCA
5951 CATCGGTATA GAATATAATC GGGGATGCCT TTATCTTGAA AAAATGCACC
6001 CGCAGCTTCG CTAGTAATCA GTAAACGCGG GAAGTGGAGT CAGGCTTTTT
6051 TTATGGAAGA GAAAATAGAC ACCAAAGTAG CCTTCTTCTA ACCTTAACGG
6101 ACCTACAGTG CAAAAAGTTA TCAAGAGACT GCATTATAGA GCGCACAAAG
6151 GAGAAAAAAA GTAATCTAAG ATGCTTTGTT AGAAAAATAG CGCTCTCGGG
6201 ATGCATTTTT GTAGAACAAA AAAGAAGTAT AGATTCTTTG TTGGTAAAAT
6251 AGCGCTCTCG CGTTGCATTT CTGTTCTGTA AAAATGCAGC TCAGATTCTT
6301 TGTTTGAAAA ATTAGCGCTC TCGCGTTGCA TTTTTGTTTT ACAAAAATGA
6351 AGCACAGATT CTTCGTTGGT AAAATAGCGC TTTCGCGTTG CATTTCTGTT
6401 CTGTAAAAAT GCAGCTCAGA TTCTTTGTTT GAAAAATTAG CGCTCTCGCG
6451 TTGCATTTTT GTTCTACAAA ATGAAGCACA GATGCTTCGT T
pBD-GAL4-Cam载体图谱质粒图谱pdf版和pBD-GAL4-Cam载体序列质粒序列等相关资料下载
pBD-GAL4-Cam质粒载体应用举例
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