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pRSET-CFP载体

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  • ¥2000
  • ZYbscience
  • 中国/美国
  • ZY35220
  • 2025年07月11日
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    • 详细信息
    • 文献和实验
    • 技术资料
    • 保存条件

      -20℃低温保存

    • 保质期

      三年

    • 英文名

      pRSET-CFP

    • 库存

      20

    • 供应商

      泽叶生物

    载体基本信息

    出品公司: ZYbscience
    载体名称: pRSET-CFP
    质粒类型: 大肠杆菌表达载体;克隆载体
    高拷贝/低拷贝: 高拷贝
    克隆方法: 多克隆位点,限制性内切酶
    启动子: T7
    载体大小: 3.6 kb
    5' 测序引物及序列: T7 Forward
    3' 测序引物及序列: --
    载体标签: His Tag (6x);Xpress Epitope Tag
    载体抗性: 氨苄青霉素(Ampicillin)
    克隆菌株: DH5alpha,TOP10F'
    表达菌株: BL21(DE3)系列,推荐BL21Star(DE3)
    备注: pRSET-EmGFP载体的功能是克隆荧光报告基因EmGFP或荧光蛋白EmGFP的表达和纯化,菌体裂解物在紫外灯下肉眼可见荧光。
    含 EK (Enterokinase)识别位点
    产品目录号: ZY35220
    稳定性: --
    组成型/诱导型: 诱导型(IPTG)
    病毒/非病毒: 非病毒

    载体质粒图谱和多克隆位点信息

    pRSET-CFP载体图谱



    pRSET-CFP载体特征

    载体简介

    载体序列

    LOCUS       pRSET/CFP	3600 bp 	DNA	SYN
    DEFINITION  pRSET/CFP
    ACCESSION   
    KEYWORDS    
    SOURCE      
      ORGANISM  other sequences; artificial sequences; vectors.
    FEATURES             Location/Qualifiers
         source          1..3600
                         /organism="pRSET/CFP"
                         /mol_type="other DNA"
         promoter        9..27
                         /label="T7_promoter"
         misc_feature    64..80
                         /label="T7_transl_en_RBS"
         CDS             89..928
                         /label="ORF frame 2"
                         /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDRWGSEFATMVSK
                         GEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLV
                         TTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTL
                         VNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAHFKIRHNIEDGSVQ
                         LADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDE
                         LYK*"
         misc_feature    101..118
                         /label="6xHIS"
                         /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDRWGSEFATMVSK
                         GEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLV
                         TTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTL
                         VNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAHFKIRHNIEDGSVQ
                         LADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDE
                         LYK*"
         misc_feature    121..139
                         /label="Xpress_fwd_primer"
                         /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDRWGSEFATMVSK
                         GEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLV
                         TTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTL
                         VNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAHFKIRHNIEDGSVQ
                         LADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDE
                         LYK*"
         misc_feature    122..154
                         /label="T7_leader"
                         /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDRWGSEFATMVSK
                         GEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLV
                         TTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTL
                         VNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAHFKIRHNIEDGSVQ
                         LADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDE
                         LYK*"
         misc_feature    122..154
                         /label="T7 Tag"
                         /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDRWGSEFATMVSK
                         GEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLV
                         TTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTL
                         VNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAHFKIRHNIEDGSVQ
                         LADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDE
                         LYK*"
         misc_feature    158..181
                         /label="Xpress_EK"
                         /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDRWGSEFATMVSK
                         GEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLV
                         TTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTL
                         VNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAHFKIRHNIEDGSVQ
                         LADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDE
                         LYK*"
         CDS             187..942
                         /label="ORF frame 1"
                         /translation="MGIRIRHHGEQGRGAVHRGGAHPGRAGRRRKRPQVQRVRRGRGR
                         CHLRQADPEVHLHHRQAARALAHPRDHPDLGRAVLQPLPRPHEAARLLQVRHARRLRP
                         GAYHLLQGRRQLQDPRRGEVRGRHPGEPHRAEGHRLQGGRQHPGAQAGVQLHQPQRLY
                         HRRQAEERHQGPLQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQR
                         PQREARSHGPAGVRDRRRDHSRHGRAVQVTREA*"
         gene            212..925
                         /label="ECFP"
                         /gene="ECFP"
                         /translation="MGIRIRHHGEQGRGAVHRGGAHPGRAGRRRKRPQVQRVRRGRGR
                         CHLRQADPEVHLHHRQAARALAHPRDHPDLGRAVLQPLPRPHEAARLLQVRHARRLRP
                         GAYHLLQGRRQLQDPRRGEVRGRHPGEPHRAEGHRLQGGRQHPGAQAGVQLHQPQRLY
                         HRRQAEERHQGPLQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQR
                         PQREARSHGPAGVRDRRRDHSRHGRAVQVTREA*"
         misc_feature    complement(254..275)
                         /label="EGFP_N_primer"
                         /translation="MGIRIRHHGEQGRGAVHRGGAHPGRAGRRRKRPQVQRVRRGRGR
                         CHLRQADPEVHLHHRQAARALAHPRDHPDLGRAVLQPLPRPHEAARLLQVRHARRLRP
                         GAYHLLQGRRQLQDPRRGEVRGRHPGEPHRAEGHRLQGGRQHPGAQAGVQLHQPQRLY
                         HRRQAEERHQGPLQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQR
                         PQREARSHGPAGVRDRRRDHSRHGRAVQVTREA*"
         misc_feature    392..421
                         /label="Y66W (ECFP)"
                         /translation="MGIRIRHHGEQGRGAVHRGGAHPGRAGRRRKRPQVQRVRRGRGR
                         CHLRQADPEVHLHHRQAARALAHPRDHPDLGRAVLQPLPRPHEAARLLQVRHARRLRP
                         GAYHLLQGRRQLQDPRRGEVRGRHPGEPHRAEGHRLQGGRQHPGAQAGVQLHQPQRLY
                         HRRQAEERHQGPLQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQR
                         PQREARSHGPAGVRDRRRDHSRHGRAVQVTREA*"
         misc_feature    862..883
                         /label="EGFP_C_primer"
                         /translation="MGIRIRHHGEQGRGAVHRGGAHPGRAGRRRKRPQVQRVRRGRGR
                         CHLRQADPEVHLHHRQAARALAHPRDHPDLGRAVLQPLPRPHEAARLLQVRHARRLRP
                         GAYHLLQGRRQLQDPRRGEVRGRHPGEPHRAEGHRLQGGRQHPGAQAGVQLHQPQRLY
                         HRRQAEERHQGPLQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQR
                         PQREARSHGPAGVRDRRRDHSRHGRAVQVTREA*"
         terminator      948..1076
                         /label="T7_terminator"
                         /translation="MGIRIRHHGEQGRGAVHRGGAHPGRAGRRRKRPQVQRVRRGRGR
                         CHLRQADPEVHLHHRQAARALAHPRDHPDLGRAVLQPLPRPHEAARLLQVRHARRLRP
                         GAYHLLQGRRQLQDPRRGEVRGRHPGEPHRAEGHRLQGGRQHPGAQAGVQLHQPQRLY
                         HRRQAEERHQGPLQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQR
                         PQREARSHGPAGVRDRRRDHSRHGRAVQVTREA*"
         misc_feature    complement(990..1008)
                         /label="T7_Terminal_primer"
                         /translation="MGIRIRHHGEQGRGAVHRGGAHPGRAGRRRKRPQVQRVRRGRGR
                         CHLRQADPEVHLHHRQAARALAHPRDHPDLGRAVLQPLPRPHEAARLLQVRHARRLRP
                         GAYHLLQGRRQLQDPRRGEVRGRHPGEPHRAEGHRLQGGRQHPGAQAGVQLHQPQRLY
                         HRRQAEERHQGPLQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQR
                         PQREARSHGPAGVRDRRRDHSRHGRAVQVTREA*"
         rep_origin      1165..1471
                         /label="f1_origin"
                         /translation="MGIRIRHHGEQGRGAVHRGGAHPGRAGRRRKRPQVQRVRRGRGR
                         CHLRQADPEVHLHHRQAARALAHPRDHPDLGRAVLQPLPRPHEAARLLQVRHARRLRP
                         GAYHLLQGRRQLQDPRRGEVRGRHPGEPHRAEGHRLQGGRQHPGAQAGVQLHQPQRLY
                         HRRQAEERHQGPLQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQR
                         PQREARSHGPAGVRDRRRDHSRHGRAVQVTREA*"
         promoter        1664..1692
                         /label="AmpR_promoter"
                         /translation="MGIRIRHHGEQGRGAVHRGGAHPGRAGRRRKRPQVQRVRRGRGR
                         CHLRQADPEVHLHHRQAARALAHPRDHPDLGRAVLQPLPRPHEAARLLQVRHARRLRP
                         GAYHLLQGRRQLQDPRRGEVRGRHPGEPHRAEGHRLQGGRQHPGAQAGVQLHQPQRLY
                         HRRQAEERHQGPLQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQR
                         PQREARSHGPAGVRDRRRDHSRHGRAVQVTREA*"
         gene            1734..2594
                         /label="Ampicillin"
                         /gene="Ampicillin"
                         /translation="MGIRIRHHGEQGRGAVHRGGAHPGRAGRRRKRPQVQRVRRGRGR
                         CHLRQADPEVHLHHRQAARALAHPRDHPDLGRAVLQPLPRPHEAARLLQVRHARRLRP
                         GAYHLLQGRRQLQDPRRGEVRGRHPGEPHRAEGHRLQGGRQHPGAQAGVQLHQPQRLY
                         HRRQAEERHQGPLQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQR
                         PQREARSHGPAGVRDRRRDHSRHGRAVQVTREA*"
         CDS             1734..2594
                         /label="ORF frame 3"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         rep_origin      2749..3368
                         /label="pBR322_origin"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
    ORIGIN
        1 CGCGAAATTA ATACGACTCA CTATAGGGAG ACCACAACGG TTTCCCTCTA GAAATAATTT
       61 TGTTTAACTT TAAGAAGGAG ATATACATAT GCGGGGTTCT CATCATCATC ATCATCATGG
      121 TATGGCTAGC ATGACTGGTG GACAGCAAAT GGGTCGGGAT CTGTACGACG ATGACGATAA
      181 GGATCGATGG GGATCCGAAT TCGCCACCAT GGTGAGCAAG GGCGAGGAGC TGTTCACCGG
      241 GGTGGTGCCC ATCCTGGTCG AGCTGGACGG CGACGTAAAC GGCCACAGGT TCAGCGTGTC
      301 CGGCGAGGGC GAGGGCGATG CCACCTACGG CAAGCTGACC CTGAAGTTCA TCTGCACCAC
      361 CGGCAAGCTG CCCGTGCCCT GGCCCACCCT CGTGACCACC CTGACCTGGG GCGTGCAGTG
      421 CTTCAGCCGC TACCCCGACC ACATGAAGCA GCACGACTTC TTCAAGTCCG CCATGCCCGA
      481 AGGCTACGTC CAGGAGCGTA CCATCTTCTT CAAGGACGAC GGCAACTACA AGACCCGCGC
      541 CGAGGTGAAG TTCGAGGGCG ACACCCTGGT GAACCGCATC GAGCTGAAGG GCATCGACTT
      601 CAAGGAGGAC GGCAACATCC TGGGGCACAA GCTGGAGTAC AACTACATCA GCCACAACGT
      661 CTATATCACC GCCGACAAGC AGAAGAACGG CATCAAGGCC CACTTCAAGA TCCGCCACAA
      721 CATCGAGGAC GGCAGCGTGC AGCTCGCCGA CCACTACCAG CAGAACACCC CCATCGGCGA
      781 CGGCCCCGTG CTGCTGCCCG ACAACCACTA CCTGAGCACC CAGTCCGCCC TGAGCAAAGA
      841 CCCCAACGAG AAGCGCGATC ACATGGTCCT GCTGGAGTTC GTGACCGCCG CCGGGATCAC
      901 TCTCGGCATG GACGAGCTGT ACAAGTAACT CGAGAAGCTT GATCCGGCTG CTAACAAAGC
      961 CCGAAAGGAA GCTGAGTTGG CTGCTGCCAC CGCTGAGCAA TAACTAGCAT AACCCCTTGG
     1021 GGCCTCTAAA CGGGTCTTGA GGGGTTTTTT GCTGAAAGGA GGAACTATAT CCGGATCTGG
     1081 CGTAATAGCG AAGAGGCCCG CACCGATCGC CCTTCCCAAC AGTTGCGCAG CCTGAATGGC
     1141 GAATGGGACG CGCCCTGTAG CGGCGCATTA AGCGCGGCGG GTGTGGTGGT TACGCGCAGC
     1201 GTGACCGCTA CACTTGCCAG CGCCCTAGCG CCCGCTCCTT TCGCTTTCTT CCCTTCCTTT
     1261 CTCGCCACGT TCGCCGGCTT TCCCCGTCAA GCTCTAAATC GGGGGCTCCC TTTAGGGTTC
     1321 CGATTTAGTG CTTTACGGCA CCTCGACCCC AAAAAACTTG ATTAGGGTGA TGGTTCACGT
     1381 AGTGGGCCAT CGCCCTGATA GACGGTTTTT CGCCCTTTGA CGTTGGAGTC CACGTTCTTT
     1441 AATAGTGGAC TCTTGTTCCA AACTGGAACA ACACTCAACC CTATCTCGGT CTATTCTTTT
     1501 GATTTATAAG GGATTTTGCC GATTTCGGCC TATTGGTTAA AAAATGAGCT GATTTAACAA
     1561 AAATTTAACG CGAATTTTAA CAAAATATTA ACGCTTACAA TTTAGGTGGC ACTTTTCGGG
     1621 GAAATGTGCG CGGAACCCCT ATTTGTTTAT TTTTCTAAAT ACATTCAAAT ATGTATCCGC
     1681 TCATGAGACA ATAACCCTGA TAAATGCTTC AATAATATTG AAAAAGGAAG AGTATGAGTA
     1741 TTCAACATTT CCGTGTCGCC CTTATTCCCT TTTTTGCGGC ATTTTGCCTT CCTGTTTTTG
     1801 CTCACCCAGA AACGCTGGTG AAAGTAAAAG ATGCTGAAGA TCAGTTGGGT GCACGAGTGG
     1861 GTTACATCGA ACTGGATCTC AACAGCGGTA AGATCCTTGA GAGTTTTCGC CCCGAAGAAC
     1921 GTTTTCCAAT GATGAGCACT TTTAAAGTTC TGCTATGTGG CGCGGTATTA TCCCGTATTG
     1981 ACGCCGGGCA AGAGCAACTC GGTCGCCGCA TACACTATTC TCAGAATGAC TTGGTTGAGT
     2041 ACTCACCAGT CACAGAAAAG CATCTTACGG ATGGCATGAC AGTAAGAGAA TTATGCAGTG
     2101 CTGCCATAAC CATGAGTGAT AACACTGCGG CCAACTTACT TCTGACAACG ATCGGAGGAC
     2161 CGAAGGAGCT AACCGCTTTT TTGCACAACA TGGGGGATCA TGTAACTCGC CTTGATCGTT
     2221 GGGAACCGGA GCTGAATGAA GCCATACCAA ACGACGAGCG TGACACCACG ATGCCTGTAG
     2281 CAATGGCAAC AACGTTGCGC AAACTATTAA CTGGCGAACT ACTTACTCTA GCTTCCCGGC
     2341 AACAATTAAT AGACTGGATG GAGGCGGATA AAGTTGCAGG ACCACTTCTG CGCTCGGCCC
     2401 TTCCGGCTGG CTGGTTTATT GCTGATAAAT CTGGAGCCGG TGAGCGTGGG TCTCGCGGTA
     2461 TCATTGCAGC ACTGGGGCCA GATGGTAAGC CCTCCCGTAT CGTAGTTATC TACACGACGG
     2521 GGAGTCAGGC AACTATGGAT GAACGAAATA GACAGATCGC TGAGATAGGT GCCTCACTGA
     2581 TTAAGCATTG GTAACTGTCA GACCAAGTTT ACTCATATAT ACTTTAGATT GATTTAAAAC
     2641 TTCATTTTTA ATTTAAAAGG ATCTAGGTGA AGATCCTTTT TGATAATCTC ATGACCAAAA
     2701 TCCCTTAACG TGAGTTTTCG TTCCACTGAG CGTCAGACCC CGTAGAAAAG ATCAAAGGAT
     2761 CTTCTTGAGA TCCTTTTTTT CTGCGCGTAA TCTGCTGCTT GCAAACAAAA AAACCACCGC
     2821 TACCAGCGGT GGTTTGTTTG CCGGATCAAG AGCTACCAAC TCTTTTTCCG AAGGTAACTG
     2881 GCTTCAGCAG AGCGCAGATA CCAAATACTG TTCTTCTAGT GTAGCCGTAG TTAGGCCACC
     2941 ACTTCAAGAA CTCTGTAGCA CCGCCTACAT ACCTCGCTCT GCTAATCCTG TTACCAGTGG
     3001 CTGCTGCCAG TGGCGATAAG TCGTGTCTTA CCGGGTTGGA CTCAAGACGA TAGTTACCGG
     3061 ATAAGGCGCA GCGGTCGGGC TGAACGGGGG GTTCGTGCAC ACAGCCCAGC TTGGAGCGAA
     3121 CGACCTACAC CGAACTGAGA TACCTACAGC GTGAGCTATG AGAAAGCGCC ACGCTTCCCG
     3181 AAGGGAGAAA GGCGGACAGG TATCCGGTAA GCGGCAGGGT CGGAACAGGA GAGCGCACGA
     3241 GGGAGCTTCC AGGGGGAAAC GCCTGGTATC TTTATAGTCC TGTCGGGTTT CGCCACCTCT
     3301 GACTTGAGCG TCGATTTTTG TGATGCTCGT CAGGGGGGCG GAGCCTATGG AAAAACGCCA
     3361 GCAACGCGGC CTTTTTACGG TTCCTGGCCT TTTGCTGGCC TTTTGCTCAC ATGTTCTTTC
     3421 CTGCGTTATC CCCTGATTCT GTGGATAACC GTATTACCGC CTTTGAGTGA GCTGATACCG
     3481 CTCGCCGCAG CCGAACGACC GAGCGCAGCG AGTCAGTGAG CGAGGAAGCG GAAGAGCGCC
     3541 CAATACGCAA ACCGCCTCTC CCCGCGCGTT GGCCGATTCA TTAATGCAGG ATCTCGATCC

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    图标文献和实验
    相关实验
    • 一文搞清蛋白质相互作用的研究方法 | 实验时间

      基因表达,比如转录因子,这样,这类蛋白就没办法通过酵母双杂交进行筛选和验证相互作用了(当然也可以将该蛋白换至 AD 载体上,但是依然有其局限性)。荧光共定位,Fluorescenceco-localization,在过去,这个技术一般用于细胞内辅助证明蛋白相互作用,前些年,如果是其他物种的蛋白质,你应用酵母双杂交系统验证了它们的相互作用,reviewer 可能会说你这个并不能反映原物种中的真实情况,可能是假阳性。这个时候,一些研究者就将这两个蛋白分别与不同颜色的荧光蛋白融合表达于原物种细胞当中,在高分辨显微

    • 一文搞清蛋白质相互作用的研究方法

      基因表达,比如转录因子,这样,这类蛋白就没办法通过酵母双杂交进行筛选和验证相互作用了(当然也可以将该蛋白换至 AD 载体上,但是依然有其局限性)。【二】荧光共定位,Fluorescenceco-localization,在过去,这个技术一般用于细胞内辅助证明蛋白相互作用,前些年,如果是其他物种的蛋白质,你应用酵母双杂交系统验证了它们的相互作用,reviewer 可能会说你这个并不能反映原物种中的真实情况,可能是假阳性。这个时候,一些研究者就将这两个蛋白分别与不同颜色的荧光蛋白融合表达于原物种细胞当中,在高分

    • 细胞共定位

      步骤 1. 载体的构建 pA7- CFP- AtCOP1,pA7-AtCRY1-YFP pA7- CFP-OsCOPl,pA7-OsCRY1b-YFP 中间载体pA7-CFP和pA7-YFP为本实验室构建。将AtCOPl(引物为AtCOP1XhoI5' 和AtCOP1 SpeI3')和AtCRYl(引物为AtCRY1XhoI5'和AtCCT1SpeI3')分别用高保真聚合酶(pfu ultra)进行PCR扩增,纯化PCR产物,用XhoI/SpeI双酶

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