pCMV-DsRed-Express2

价格:2000元

联系方式:I47-825O-882O

买家导航

载体基本信息

出品公司: Clontech
载体名称: pCMV-DsRed-Express2
质粒类型: 慢病毒载体;荧光报告载体
高拷贝/低拷贝: 高拷贝
克隆方法: 限制性内切酶,多克隆位点
启动子: CMV
载体大小: 4638 bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: --
载体抗性: 卡那霉素
筛选标记: 新霉素(Neomycin)
克隆菌株: DH5α, HB101
宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
备注: pCMV-DsRed-Express2组成型表达DsRed-Express2;
在DsRed-Express2的上下游留有酶切位点,便于转移到其它载体上去;
DsRed-Express2是DsRed的突变体,与DsRed相比,DsRed-Express2细胞毒性低,尤其适用于原代细胞和干细胞的荧光标记。
产品目录号: 632539
稳定性: 稳表达 或 瞬表达
组成型/诱导型: 组成型
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息

pCMV-DsRed-Express2载体图谱



pCMV-DsRed-Express2 载体特征

载体简介

pCMV-DsRed-Express2载体描述

pCMV DsRed-Express2 is a mammalian expression vector designed to be used for whole cell labeling or as a marker of cotransfection. In mammalian cells, the vector constitutively expresses DsRed-Express2, a variant of the Discosoma sp. red fluorescent protein, DsRed (1). DsRed-Express2 retains the fast maturation and high photostability characteristic of its predecessor, DsRed-Express (2), and has been engineered (through additional amino acid substitutions) for increased solubility (3). Although it most likely forms the same tetrameric structure as wild-type DsRed, DsRed-Express2 displays a greatly reduced tendency to aggregate, resulting in reduced cyto- and phototoxicity, and making DsRed-Express2 much better suited for in vivo applications involving sensitive cells, such as primary or stem cells. In fact, DsRed-Express2 has been shown to be the best red fluorescent protein for whole cell labeling applications (3). DsRed-Express2 also exhibits extremely low residual green fluorescence, which allows cells expressing the protein to be effectively separated from other fluorescently labeled cell populations by flow cytometry. The DsRed-Express2 gene is positioned just downstream of the constitutively active human cytomegalovirus immediate early promoter (PCMV IE). As a result, mammalian cells transfected with this vector will constitutively express the red fluorescent protein. A Kozak consensus sequence (4) has been placed immediately upstream of the DsRed-Express2 coding sequence to enhance translational efficiency in eukaryotic cells. SV40 polyadenylation signals downstream of the DsRed-Express2 gene direct proper processing of the 3' end of the DsRed-Express2 mRNA. The vector backbone contains an SV40 origin for replication in mammalian cells expressing the SV40 large T antigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production. A neomycin resistance cassette (Neor) allows stably transfected eukaryotic cells to be selected using G418 (5). This cassette consists of the SV40 early promoter, a Tn5 kanamycin/neomycin resistance gene, and herpes simplex virus thymidine kinase (HSV TK) polyadenylation signals. A bacterial promoter upstream of the cassette expresses kanamycin resistance in E. coli.

pCMV DsRed-Express2 can be used for whole cell labeling or as a cotransfection marker. The vector can be transfected into mammalian cells using any standard transfection method. Cells expressing DsRed-Express2 (excitation and emission maxima: 554 nm and 591 nm, respectively) can be detected by fluorescence microscopy or flow cytometry 8-12 hours after transfection. If required, stable transfectants can be selected using G418. After cotransfection with pCMV DsRed-Express2 and an expression construct of interest, cells can also be sorted by flow cytometry to enrich for transfected cells.

Propagation in E. coli
 Recommended host strain: DH5α, HB101, and other general purpose strains. Single-stranded DNA production
requires a host containing an F plasmid such as JM109 or XL1-Blue.
 Selectable marker: plasmid confers resistance to kanamycin (50 μg/ml) in E. coli hosts.
 E. coli replication origin: pUC
 Copy number: high
 Plasmid incompatibility group: pMB1/ColE1

Excitation and emission maxima of DsRed-Express2
 Excitation maximum = 554 nm
 Emission maximum = 591 nm

References
1. Matz, M. V. et al. (1999) Nat. Biotechnol. 17(10):969-973.
2. Bevis, B. J. & Glick, B. S. (2002) Nat. Biotechnol. 20(1):83–87. Erratum in Nat. Biotechnol. (2002) 20(11):1159
3. Strack, R. L. et al. (2008) Nat. Methods 5(11):955–957.
4. Kozak, M. (1987) Nucleic Acids Res. 15(20): 8125-8148
5. Gorman, C. (1985) In DNA Cloning: A Practical Approach, Vol. II. Ed. D. M. Glover. (IRL Press, Oxford, U.K.), pp. 143–190.

载体序列

LOCUS       pCMV DsRed-Express2   4638 bp    DNA   circular             8-NOV-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 632539
FEATURES             Location/Qualifiers
     promoter        1..589
                     /label=CMV\IE\Promoter
                     /note="Human cytomegalovirus (CMV) immediate early promoter"
     CDS             2534..3328
                     /label=Kan(R)/Neo(R)
     rep_origin      3913..4556
                     /label=pUC\ori
     CDS             628..1302
                     /label=DsRed-Express2
BASE COUNT     1102 a      1252 c      1229 g      1055 t 
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 
      601 ccggactcag atccaccggt cgccaccatg gatagcactg agaacgtcat caagcccttc 
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc 
      721 gagggcgagg gcaagcccta cgagggcacc cagaccgcca agctgcaggt gaccaagggc 
      781 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg 
      841 tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc 
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc 
      961 tccctgcagg acggcacctt catctaccac gtgaagttca tcggcgtgaa cttcccctcc 
     1021 gacggccccg taatgcagaa gaagactctg ggctgggagc cctccaccga gcgcctgtac 
     1081 ccccgcgacg gcgtgctgaa gggcgagatc cacaaggcgc tgaagctgaa gggcggcggc 
     1141 cactacctgg tggagttcaa gtcaatctac atggccaaga agcccgtgaa gctgcccggc 
     1201 tactactacg tggactccaa gctggacatc acctcccaca acgaggacta caccgtggtg 
     1261 gagcagtacg agcgcgccga ggcccgccac cacctgttcc agtagcggcc gcgactctag 
     1321 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac 
     1381 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca 
     1441 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 
     1501 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt 
     1561 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa 
     1621 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt 
     1681 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa 
     1741 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag 
     1801 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt 
     1861 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg 
     1921 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc 
     1981 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg 
     2041 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 
     2101 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag 
     2161 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt 
     2221 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca 
     2281 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta 
     2341 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 
     2401 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag 
     2461 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg 
     2521 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 
     2581 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 
     2641 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 
     2701 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 
     2761 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 
     2821 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 
     2881 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 
     2941 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 
     3001 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 
     3061 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 
     3121 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 
     3181 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 
     3241 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 
     3301 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa 
     3361 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 
     3421 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 
     3481 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg 
     3541 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg 
     3601 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga 
     3661 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg 
     3721 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt 
     3781 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 
     3841 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga 
     3901 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 
     3961 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 
     4021 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 
     4081 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 
     4141 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 
     4201 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 
     4261 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 
     4321 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 
     4381 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 
     4441 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 
     4501 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 
     4561 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 
     4621 cgtattaccg ccatgcat 
//

载体质粒图谱pdf版和相关资料下载

载体应用举例

分享到:

全部载体分类

相关载体

相关文章

相关问答

Copyright © 20012-2013 BIOFENG. 生物风 版权所有 Powered by Biofeng