pCold IV

价格:1500元

联系方式:I47-825O-882O

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pColdIV载体基本信息

出品公司: Takara
载体名称: pCold IV, pColdIV, pCold-IV
质粒类型: 大肠杆菌蛋白表达载体
表达水平:
启动子: CSPA
克隆方法: 多克隆位点,限制性内切酶
载体大小: 4359 bp
5' 测序引物及序列: pCold-F: 5'-ACGCCATATCGCCGAAAGG-3'
3' 测序引物及序列: pCold-R: 5'-GGCAGGGATCTTAGATTCTG-3'
载体标签: /
载体抗性: Ampicillin 氨苄
备注:
温度诱导,冷休克蛋白表达载体
产品目录号: 3364
稳定性: 瞬时表达 Transient
组成型: 诱导表达
病毒/非病毒: 非病毒

pColdIV载体质粒图谱和多克隆位点信息

pCold IV载体图谱

pCold IV多克隆位点

pColdIV载体简介

阐明蛋白质的结构和功能是后基因组研究的一个重要课题。高效率的蛋白质表达系统是研究这些课题的不可缺少的基本技术。大肠杆菌作为宿主的表达系统中被广泛 使用于生产重组蛋白。大肠杆菌表达系统具有良好的优势,方法简单,成本低。然而,对于某些基因,蛋白表达很困难,表达的蛋白质不溶。

为了解决这些问题,TAKARA与Masayori Inouye教授(美国新泽西医学和牙科大学)进行联合研究,以制定一个高效基于低温冷休克蛋白基因的大肠杆菌表达载体。pCold DNA系列产品具有上述的优点,并提供了一个重要功能和结构的分析以及其他蛋白质研究领域的工具。

<产品概况和特点>
当培养的大肠杆菌的温度充分降低,几乎暂时停止生长大部分的蛋白表达减少,而此时一组蛋白质称为“冷休克蛋白”被专门诱导表达。

冷休克表达载体,pCold I-IV,是利用冷休克基因CSPA启动子设计高效率蛋白质表达载体。 CSPA的启动子的下游处,插入了lac操纵子,以便严格控制表达。此外,5'非翻译区(5'UTR),翻译增强元件(TEE),His标签序列,Xa因 子切割位点和多克隆位点(MCS)位于在CSPA启动子的下游的。由于该产品采用了大肠杆菌的启动子,大多数大肠杆菌菌株可被用作表达宿主。有四种类型的 pCold载体,他们的下列原件的位置稍有不同,TEE,His标签序列和Factor XA酶切位点。

pColdIV载体序列

  
LOCUS       pCold\IV                4359 bp    DNA     circular SYN 10-AUG-2004
DEFINITION  Expression vector pColdIV DNA, complete sequence.
ACCESSION   AB186391
VERSION     AB186391.1  GI:51090256
KEYWORDS    .
SOURCE      Expression vector pColdIV
  ORGANISM  Expression vector pColdIV
            other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     VNTAUTHORNAME|biofeng.com|
FEATURES             Location/Qualifiers
     source          1..4359
                     /organism="Expression vector pColdIV"
                     /mol_type="other DNA"
                     /db_xref="taxon:289232"
                     /lab_host="Escherichia coli"
                     /note="expresses an open reading frame directly. This
                     plasmid is derived from pUC118."
     gene            15..464
                     /gene="cspA"
     promoter        15..81
                     /gene="cspA"
     misc_feature    82..121
                     /gene="cspA"
                     /note="synthetic lac operator sequence"
     5'UTR           122..255
                     /gene="cspA"
                     /note="modified cspA 5'UTR"
     misc_feature    253..312
                     /gene="cspA"
                     /note="MCS = Multiple cloning sites contain the follow
                     restriction sites: NdeI, SacI, KpnI, XhoI, BamHI, EcoRI,
                     HindIII, SalI, PstI, XbaI"
     3'UTR           320..464
                     /gene="cspA"
                     /note="cspA 3'UTR"
     misc_feature    complement(663..1136)
                     /note="M13 intergenic region"
     gene            1323..2183
                     /gene="Amp"
     CDS             1323..2183
                     /gene="Amp"
                     /codon_start=1
                     /transl_table=11
                     /product="beta-lactamase"
                     /protein_id="BAD35140.1"
                     /db_xref="GI:51090257"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     rep_origin      2343..2957
                     /note="ColE1 origin"
     gene            complement(3114..4196)
                     /gene="lacI"
     CDS             complement(3114..4196)
                     /gene="lacI"
                     /codon_start=1
                     /transl_table=11
                     /product="lactose operon repressor"
                     /protein_id="BAD35141.1"
                     /db_xref="GI:51090258"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
                     LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
                     GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
                     DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
                     EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
                     QTASPRALADSLMQLARQVSRLESGQ"
ORIGIN      
        1 aaggaatggt gtggccgatt aatcataaat atgaaaaata attgttgcat cacccgccaa
       61 tgcgtggctt aatgcacatc aaattgtgag cggataacaa tttgatgtgc tagcgcatat
      121 ccagtgtagt aaggcaagtc ccttcaagag ttatcgttga tacccctcgt agtgcacatt
      181 cctttaacgc ttcaaaatct gtaaagcacg ccatatcgcc gaaaggcaca cttaattatt
      241 aagaggtaat accatatgga gctcggtacc ctcgagggat ccgaattcaa gcttgtcgac
      301 ctgcagtcta gataggtaat ctctgcttaa aagcacagaa tctaagatcc ctgccatttg
      361 gcggggattt ttttatttgt tttcaggaaa taaataatcg atcgcgtaat aaaatctatt
      421 attatttttg tgaagaataa atttgggtgc aatgagaatg cgcaggccct ttcgtctcgc
      481 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
      541 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      601 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      661 taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      721 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
      781 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      841 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      901 caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      961 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     1021 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     1081 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg
     1141 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg tcaggtggca
     1201 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1261 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1321 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1381 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1441 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1501 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1561 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1621 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1681 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1741 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1801 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1861 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1921 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1981 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2041 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2101 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2161 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2221 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2281 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2341 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2401 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2461 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2521 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2581 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2641 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2701 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2761 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2821 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2881 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2941 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3001 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     3061 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     3121 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3181 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     3241 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     3301 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     3361 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     3421 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     3481 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     3541 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     3601 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     3661 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     3721 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     3781 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     3841 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     3901 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     3961 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     4021 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     4081 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     4141 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     4201 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     4261 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
     4321 gcccagtagt aggttgaggc cgttgagcac cgccgccgc
//

pColdIV载体质粒图谱pdf版和相关资料下载

pColdIV载体应用举例

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