pTrcHis A编号 载体名称北京华越洋生物VECT4450 pTrcHis ApTrcHisA载体基本信息别名: pTrcHisA, p TrcHis A质粒类型: 大肠杆菌蛋白表达载体表达水平: 高拷贝启动子: Trc克隆方法: 多克隆位点，限制性内切酶载体大小: 4414 b p5' 测序引物及序列: pTrcHis-‐F: g aggtatatattaatgtatcg3' 测序引物及序列: pTrcHis-‐R:GATTTAATCTGTATCAGG 载体标签: N-‐His, N-‐EK载体抗性: Ampicillin备注: Hosts: E.coli NM522, E.coli INValphaF'. Related vectors: pSE420.稳定性: 瞬时表达 Transient 组成型: 组成型 Constitutive 病毒/非病毒: 非病毒pTrcHisA载体质粒图谱和多克隆位点信息pTrcHisA载体简介pTrcHis载体是来由pBR322质粒衍生而来的表达载体,设计用来在大肠杆菌中进行重组蛋白的高效表达和纯化。
载体上的Trc启动子和rrnB抗终止区使得高水平蛋白表达成为可能.Trc启动子由Trp启动子的-‐35区域和Lac启动子的-‐10区域共同组成。
pTrcHis载体同时包含有一个LacIq 基因，该基因编码了Lac启动子的抑制蛋白。
LacIq基因的存在使得宿主菌是LacIq+还是LacIq-‐都可以高效抑制插入基因的转录。
进行蛋白表达时，大肠杆菌需要生长到对数生长期，然后加入1mM I PTG，去除阻遏蛋白，诱导表达。
载体上的迷你顺反子结构能够增强插入基因的转录水平，从而高水平表达目的蛋白。
DNA插入片段位于载体表达片段下游，在其上游有一个N-‐端融合表达多肽序列，该序列包含有ATG起始密码子，6xHis标签，T7噬菌体10号基因的翻译增强子，X p ress 抗原表位，肠激酶酶切识别位点。
pTrcHisA载体序列ORIGIN1 GTTTGACAGC TTATCATCGA CTGCACGGTG CACCAATGCT TCTGGCGTCA GGCAGCCATC 61 GGAAGCTGTG GTATGGCTGT GCAGGTCGTA AATCACTGCA TAATTCGTGT CGCTCAAGGC 121 GCACTCCCGT TCTGGATAAT GTTTTTTGCG CCGACATCAT AACGGTTCTG GCAAATATTC 181 TGAAATGAGC TGTTGACAAT TAATCATCCG GCTCGTATAA TGTGTGGAAT TGTGAGCGGA 241 TAACAATTTC ACACAGGAAA CAGCGCCGCT GAGAAAAAGC GAAGCGGCAC TGCTCTTTAA 301 CAATTTATCA GACAATCTGT GTGGGCACTC GACCGGAATT ATCGATTAAC TTTATTATTA 361 AAAATTAAAG AGGTATATAT TAATGTATCG ATTAAATAAG GAGGAATAAA CCATGGGGGG 421 TTCTCATCAT CATCATCATC ATGGTATGGC TAGCATGACT GGTGGACAGC AAATGGGTCG481 GGATCTGTAC GACGATGACG ATAAGGATCG ATGGGGATCC GAGCTCGAGA TCTGCAGCTG 541 GTACCATATG GGAATTCGAA GCTTGGCTGT TTTGGCGGAT GAGAGAAGAT TTTCAGCCTG 601 ATACAGATTA AATCAGAACG CAGAAGCGGT CTGATAAAAC AGAATTTGCC TGGCGGCAGT 661 AGCGCGGTGG TCCCACCTGA CCCCATGCCG AACTCAGAAG TGAAACGCCG TAGCGCCGAT 721 GGTAGTGTGG GGTCTCCCCA TGCGAGAGTA GGGAACTGCC AGGCATCAAA TAAAACGAAA 781 GGCTCAGTCG AAAGACTGGG CCTTTCGTTT TATCTGTTGT TTGTCGGTGA ACGCTCTCCT 841 GAGTAGGACA AATCCGCCGG GAGCGGATTT GAACGTTGCG AAGCAACGGC CCGGAGGGTG 901 GCGGGCAGGA CGCCCGCCAT AAACTGCCAG GCATCAAATT AAGCAGAAGG CCATCCTGAC 961 GGATGGCCTT TTTGCGTTTC TACAAACTCT TTTTGTTTAT TTTTCTAAAT ACATTCAAAT 1021 ATGTATCCGC TCATGAGACA ATAACCCTGA TAAATGCTTC AATAATATTG AAAAAGGAAG 1081 AGTATGAGTA TTCAACATTT CCGTGTCGCC CTTATTCCCT TTTTTGCGGC ATTTTGCCTT 1141 CCTGTTTTTG CTCACCCAGA AACGCTGGTG AAAGTAAAAG ATGCTGAAGA TCAGTTGGGT 1201 GCACGAGTGG GTTACATCGA ACTGGATCTC AACAGCGGTA AGATCCTTGA GAGTTTTCGC 1261 CCCGAAGAAC GTTTTCCAAT GATGAGCACT TTTAAAGTTC TGCTATGTGG CGCGGTATTA 1321 TCCCGTGTTG ACGCCGGGCA AGAGCAACTC GGTCGCCGCA TACACTATTC TCAGAATGAC 1381 TTGGTTGAGT ACTCACCAGT CACAGAAAAG CATCTTACGG ATGGCATGAC AGTAAGAGAA 1441 TTATGCAGTG CTGCCATAAC CATGAGTGAT AACACTGCGG CCAACTTACT TCTGACAACG 1501 ATCGGAGGAC CGAAGGAGCT AACCGCTTTT TTGCACAACA TGGGGGATCA TGTAACTCGC 1561 CTTGATCGTT GGGAACCGGA GCTGAATGAA GCCATACCAA ACGACGAGCG TGACACCACG 1621 ATGCCTGTAG CAATGGCAAC AACGTTGCGC AAACTATTAA CTGGCGAACT ACTTACTCTA 1681 GCTTCCCGGC AACAATTAAT AGACTGGATG GAGGCGGATA AAGTTGCAGG ACCACTTCTG 1741 CGCTCGGCCC TTCCGGCTGG CTGGTTTATT GCTGATAAAT CTGGAGCCGG TGAGCGTGGG 1801 TCTCGCGGTA TCATTGCAGC ACTGGGGCCA GATGGTAAGC CCTCCCGTAT CGTAGTTATC 1861 TACACGACGG GGAGTCAGGC AACTATGGAT GAACGAAATA GACAGATCGC TGAGATAGGT 1921 GCCTCACTGA TTAAGCATTG GTAACTGTCA GACCAAGTTT ACTCATATAT ACTTTAGATT 1981 GATTTAAAAC TTCATTTTTA ATTTAAAAGG ATCTAGGTGA AGATCCTTTT TGATAATCTC 2041 ATGACCAAAA TCCCTTAACG TGAGTTTTCG TTCCACTGAG CGTCAGACCC CGTAGAAAAG 2101 ATCAAAGGAT CTTCTTGAGA TCCTTTTTTT CTGCGCGTAA TCTGCTGCTT GCAAACAAAA 2161 AAACCACCGC TACCAGCGGT GGTTTGTTTG CCGGATCAAG AGCTACCAAC TCTTTTTCCG 2221 AAGGTAACTG GCTTCAGCAG AGCGCAGATA CCAAATACTG TCCTTCTAGT GTAGCCGTAG 2281 TTAGGCCACC ACTTCAAGAA CTCTGTAGCA CCGCCTACAT ACCTCGCTCT GCTAATCCTG 2341 TTACCAGTGG CTGCTGCCAG TGGCGATAAG TCGTGTCTTA CCGGGTTGGA CTCAAGACGA 2401 TAGTTACCGG ATAAGGCGCA GCGGTCGGGC TGAACGGGGG GTTCGTGCAC ACAGCCCAGC 2461 TTGGAGCGAA CGACCTACAC CGAACTGAGA TACCTACAGC GTGAGCTATG AGAAAGCGCC 2521 ACGCTTCCCG AAGGGAGAAA GGCGGACAGG TATCCGGTAA GCGGCAGGGT CGGAACAGGA 2581 GAGCGCACGA GGGAGCTTCC AGGGGGAAAC GCCTGGTATC TTTATAGTCC TGTCGGGTTT 2641 CGCCACCTCT GACTTGAGCG TCGATTTTTG TGATGCTCGT CAGGGGGGCG GAGCCTATGG 2701 AAAAACGCCA GCAACGCGGC CTTTTTACGG TTCCTGGCCT TTTGCTGGCC TTTTGCTCAC 2761 ATGTTCTTTC CTGCGTTATC CCCTGATTCT GTGGATAACC GTATTACCGC CTTTGAGTGA 2821 GCTGATACCG CTCGCCGCAG CCGAACGACC GAGCGCAGCG AGTCAGTGAG CGAGGAAGCG 2881 GAAGAGCGCC TGATGCGGTA TTTTCTCCTT ACGCATCTGT GCGGTATTTC ACACCGCATA 2941 TGGTGCACTC TCAGTACAAT CTGCTCTGAT GCCGCATAGT TAAGCCAGTA TACACTCCGC 3001 TATCGCTACG TGACTGGGTC ATGGCTGCGC CCCGACACCC GCCAACACCC GCTGACGCGC 3061 CCTGACGGGC TTGTCTGCTC CCGGCATCCG CTTACAGACA AGCTGTGACC GTCTCCGGGA3121 GCTGCATGTG TCAGAGGTTT TCACCGTCAT CACCGAAACG CGCGAGGCAG CAGATCAATT 3181 CGCGCGCGAA GGCGAAGCGG CATGCATTTA CGTTGACACC ATCGAATGGC GCAAAACCTT 3241 TCGCGGTATG GCATGATAGC GCCCGGAAGA GAGTCAATTC AGGGTGGTGA ATGTGAAACC 3301 AGTAACGTTA TACGATGTCG CAGAGTATGC CGGTGTCTCT TATCAGACCG TTTCCCGCGT 3361 GGTGAACCAG GCCAGCCACG TTTCTGCGAA AACGCGGGAA AAAGTGGAAG CGGCGATGGC 3421 GGAGCTGAAT TACATTCCCA ACCGCGTGGC ACAACAACTG GCGGGCAAAC AGTCGTTGCT 3481 GATTGGCGTT GCCACCTCCA GTCTGGCCCT GCACGCGCCG TCGCAAATTG TCGCGGCGAT 3541 TAAATCTCGC GCCGATCAAC TGGGTGCCAG CGTGGTGGTG TCGATGGTAG AACGAAGCGG 3601 CGTCGAAGCC TGTAAAGCGG CGGTGCACAA TCTTCTCGCG CAACGCGTCA GTGGGCTGAT 3661 CATTAACTAT CCGCTGGATG ACCAGGATGC CATTGCTGTG GAAGCTGCCT GCACTAATGT 3721 TCCGGCGTTA TTTCTTGATG TCTCTGACCA GACACCCATC AACAGTATTA TTTTCTCCCA 3781 TGAAGACGGT ACGCGACTGG GCGTGGAGCA TCTGGTCGCA TTGGGTCACC AGCAAATCGC 3841 GCTGTTAGCG GGCCCATTAA GTTCTGTCTC GGCGCGTCTG CGTCTGGCTG GCTGGCATAA 3901 ATATCTCACT CGCAATCAAA TTCAGCCGAT AGCGGAACGG GAAGGCGACT GGAGTGCCAT 3961 GTCCGGTTTT CAACAAACCA TGCAAATGCT GAATGAGGGC ATCGTTCCCA CTGCGATGCT 4021 GGTTGCCAAC GATCAGATGG CGCTGGGCGC AATGCGCGCC ATTACCGAGT CCGGGCTGCG 4081 CGTTGGTGCG GATATCTCGG TAGTGGGATA CGACGATACC GAAGACAGCT CATGTTATAT 4141 CCCGCCGTCA ACCACCATCA AACAGGATTT TCGCCTGCTG GGGCAAACCA GCGTGGACCG 4201 CTTGCTGCAA CTCTCTCAGG GCCAGGCGGT GAAGGGCAAT CAGCTGTTGC CCGTCTCACT 4261 GGTGAAAAGA AAAACCACCC TGGCGCCCAA TACGCAAACC GCCTCTCCCC GCGCGTTGGC 4321 CGATTCATTA ATGCAGCTGG CACGACAGGT TTCCCGACTG GAAAGCGGGC AGTGAGCGCA 4381 ACGCAATTAA TGTGAGTTAG CGCGAATTGA TCTG//其他大肠杆菌表达载体：pBV221 ptdTomato pET-52b(+) pAmCyanpDsRed-Express2 pBV220 pCold-GST pColdS-SUMOpCold TF pCold IV pCold III pCold IIpCold I pE-SUMO pCold-ProS2 pBAD102/D-TOPOpBAD202/D-TOPO pACYC184 pBAD/Thio-TOPO pBad/Myc-His CpBad/Myc-His B pBad/Myc-His A pBad/His C pBad/His BpBad/His A pBAD-TOPO pET-23b(+) pET-23a(+)pET-23c(+) pET-23(+) pET-12b(+) pET-12c(+)pET-12a(+) pET-11b(+) pET-11a(+) pET-11c(+)pBad24 pQE-82L pQE-81L pQE-80LpQE-32 pQE-9 pQE-16 pQE-31pQE-60 pQE-70 pQE-40 pET-51b(+)pET-50b(+) pET-49b(+) pET-48b(+) pET-47b(+)pET-26b(+) pET-32a(+) pET-21b(+) pET-22b(+)pET-14b pET-16b pET-15b pET-19bpET-20b(+) pET-21d(+) pET-21c(+) pET-21b(+)pET-21a(+) pET-24a(+) pET-24d(+) pET-25b(+)pET-27b(+) pET-28a(+) pET-30a(+) pET-42a(+)pET-43.1c(+) pET-43.1b(+) pET-43.1a(+) pET-44a(+)pET-44c(+) pET-46 EK/LIC pET-37b(+) pTrcHis2 C pTrcHis2 B pTrcHis2 A pET303/CT-His pET302/NT-His pRSET-CFP pRSET-EmGFP pRSET-BFP pGFPuvpET300/NT-DEST pET301/CT-DEST pGEM-T pBad43pGEX-4T-3 pGEX-5X-2 pBlueScript SK(+) pG-Tf2pG-KJE8 pGro7 pET-SUMO pSE380pET-17b pET102/D-TOPO pCDFDuet-1 pMAL-p5xpTf16 pET-28c(+) pBluescript II SK(+) pET-30b(+) pSUMO pProEX HTc pProEX HTb pProEX HTa pKD3 pKD13 pKD46 pTYB1pTYB2 pTWIN2 pBluescript II KS(-) pTYB12pMAL-p5e pACYCDuet-1 pEGM-11ZF(+) pEGM-7ZF(+) PinPoint Xa-3 PinPoint Xa-2 PinPoint Xa-1 pSP73pSP64 pTWIN1 pTYB11 pTXB1pET-5b(+) pBad/gIII C pBad/gIII B pBad/gIII A pET-5a(+) pMal-p4X pMal-p2G pkk223-3pkk232-8 pCYB1 pEZZ18 pBAD18pMAL-c5x pMal-p2E pMal-p2X pET-44 EK/LIC pET-43.1 EK/LIC pET-41 EK/LIC pMal-c4X pTrcHis BpET-31b(+) pET-3b(+) pET-41a(+) pGEX-3XpGEX-4T-2 pETDuet-1 pGEX-4T-1 pTrc99apET-28b(+) pET-His pALEX a,b,c pACYC177pBR322 pKD4 pKD20 pMXB10pEcoli-6xHN-GFPuv pKJE7 pRSET B pGEX-KGpGEX-2T pRSFDuet-1 pCOLADuet-1 pTrcHis C pTrcHis A pET-41b(+) pET-42b(+) pET-3a(+) pGEX-6P-3 pGEX-6P-2 pGEX-6P-1 pGEX-5X-3 pGEX-5X-1 pGEX-2TK pRSET A pMal-c2GpMal-c2E pMal-c2X pRSET C pQE-30pET-45b(+) pET-44b(+) pET-42c(+) pET-41c(+) pET-40b(+) pET-33b(+) pET-39b(+) pET-32 EK/LIC pET-32 Xa/LIC pET-32c(+) pET-32b(+) pET-30 Xa/LIC pET-30 EK/LIC pET-30c(+) pET-29c(+) pET-29b(+) pET-29a(+) pET-24c(+) pET-24b(+) pET-24(+)pET-23d(+) pET-11d(+) pBad33。