| 出品公司: | Clontech |
|---|---|
| 载体名称: | pCMV-p53 、 p53 Dominant-Negative Vector |
| 质粒类型: | 信号通路分析载体；显性抑制载体 |
| 克隆方法: | -- |
| 启动子: | CMV |
| 载体大小: | 5167 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: | Neomycin |
| 克隆菌株: | DH5a 或 HB101 |
| 宿主细胞（系）: | -- |
| 备注: |
pCMV-p53载体是信号通路分析载体； p53是信号转导蛋白，p53的突变体是显性负性突变体，抑制信号转导过程。 |
| 产品目录号: | 632420 |
| 稳定性: | 瞬表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
p53
The p53 Dominant-Negative Vector Set lets you study p53 and related signaling pathways. These vectors constitutively express high levels of wild-type p53 or its dominant-negative mutant. To determine the effects of p53 expression on a particular cell line, simply transfect with the vector containing the gene for wild-type p53, and examine morphology or look for changes in the expression of other pathway proteins.
载体描述
The p53 Dominant-Negative Vector Set is a convenient tool for monitoring p53-mediated signal transduction pathways. This vector set consists of two vectors, pCMV-p53 and pCMV-p53mt135. pCMV-p53 expresses the wild-type p53 tumor suppression protein and pCMV-p53mt135 expresses a dominant-negative mutant. Both proteins are expressed at high levels from the constitutive CMV promoter. The p53 gene and p53mt135 gene differ by a G to A conversion at nucleotide 1017.
The SV40 polyadenylation sequence directs proper processing of the 3' end of the p53 or p53mt135 mRNAs. The vector backbone contains an SV40 origin for replication in mammalian cells expressing the SV40 T antigen. A neomycin-resistance cassette (Neor)—consisting of the SV40 early promoter, the Tn5 neomycin/kanamycin resistance gene, and polyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene—allows kanamycin selection in E. coli and neomycin selection in eukaryotic cells. The vector backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.
使用
pCMV-p53 is used to examine the physical interactions between p53 and other proteins, to induce p53-mediated cell cycle arrest, or to study p53-induced gene expression. pCMV-p53mt135 expresses the p53mt135 mutant, which because of a conformational change, can no longer interact with p53-binding sites. When p53mt135 and p53 are co-expressed, they form a mixed tetramer that is unable to interact with p53-binding sites; therefore, the downstream effects of p53 are blocked (1, 2).
Both vectors can be transfected into mammalian cells using any standard method. Stable transformants can be selected using G418 (3).
Propagation in E. coli
Suitable host strains: DH5a HB101 and other general purpose strains. Single-stranded DNA production requires
a host containing an F plasmid such as JM101 or XL1-Blue.
Selectable marker: plasmid confers resistance to kanamycin (50μg/ml) in E. coli hosts.
E. coli replication origin: pUC
Copy number: ~500
Plasmid incompatibility group: pMB1/ColE1
载体序列
LOCUS pCMV-p53 5167 bp DNA circular 23-NOV-2009
COMMENT Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT Cat. No. 631922
FEATURES Location/Qualifiers
misc_feature 1..589
/label=CMV\IE\Promoter
misc_feature 613..1791
/label=p53
misc_feature 1986..1991
/label=SV40\polyA\signal
misc_feature 2016..2020
/label=SV40\polyA\signal
misc_feature 2083..2538
/label=f1\origin
misc_feature 2600..2635
/label=Bacterial\Promoter
misc_feature 2879..3014
/label=SV40\origin
scRNA 2712..2941
/label=SV40\early\Promoter
misc_feature 3063..3857
/label=Kan(R)/Neo(R)
misc_feature 4093..4098
/label=HSV\TK\polyA\signal
misc_feature 4106..4111
/label=HSV\TK\polyA\signal
misc_feature 4442..5085
/label=pUC\origin
BASE COUNT 1240 a 1395 c 1339 g 1193 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 gctagatctc
601 gagctcaagc ttatggagga gccgcagtca gatcctagcg tcgagccccc tctgagtcag
661 gaaacatttt cagacctatg gaaactactt cctgaaaaca acgttctgtc ccccttgccg
721 tcccaagcaa tggatgattt gatgctgtcc ccggacgata ttgaacaatg gttcactgaa
781 gacccaggtc cagatgaagc tcccagaatg ccagaggctg ctccccccgt ggcccctgca
841 ccagcagctc ctacaccggc ggcccctgca ccagccccct cctggcccct gtcatcttct
901 gtcccttccc agaaaaccta ccagggcagc tacggtttcc gtctgggctt cttgcattct
961 gggacagcca agtctgtgac ttgcacgtac tcccctgccc tcaacaagat gttttgccaa
1021 ctggccaaga cctgccctgt gcagctgtgg gttgattcca cacccccgcc cggcacccgc
1081 gtccgcgcca tggccatcta caagcagtca cagcacatga cggaggttgt gaggcgctgc
1141 ccccaccatg agcgctgctc agatagcgat ggtctggccc ctcctcagca tcttatccga
1201 gtggaaggaa atttgcgtgt ggagtatttg gatgacagaa acacttttcg acatagtgtg
1261 gtggtgccct atgagccgcc tgaggttggc tctgactgta ccaccatcca ctacaactac
1321 atgtgtaaca gttcctgcat gggcggcatg aaccggaggc ccatcctcac catcatcaca
1381 ctggaagact ccagtggtaa tctactggga cggaacagct ttgaggtgcg tgtttgtgcc
1441 tgtcctggga gagaccggcg cacagaggaa gagaatctcc gcaagaaagg ggagcctcac
1501 cacgagctgc ccccagggag cactaagcga gcactgccca acaacaccag ctcctctccc
1561 cagccaaaga agaaaccact ggatggagaa tatttcaccc ttcagatccg tgggcgtgag
1621 cgcttcgaga tgttccgaga gctgaatgag gccttggaac tcaaggatgc ccaggctggg
1681 aaggagccag gggggagcag ggctcactcc agccacctga agtccaaaaa gggtcagtct
1741 acctcccgcc ataaaaaact catgttcaag acagaagggc ctgactcaga ctgagaattc
1801 tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag
1861 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
1921 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
1981 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
2041 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
2101 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
2161 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
2221 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
2281 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
2341 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
2401 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
2461 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
2521 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
2581 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
2641 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
2701 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
2761 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
2821 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
2881 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
2941 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
3001 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
3061 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
3121 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
3181 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
3241 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
3301 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
3361 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
3421 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
3481 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
3541 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
3601 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
3661 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
3721 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
3781 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
3841 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
3901 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
3961 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
4021 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
4081 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
4141 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
4201 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
4261 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
4321 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
4381 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
4441 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
4501 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
4561 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
4621 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
4681 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
4741 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
4801 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
4861 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
4921 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
4981 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
5041 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
5101 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
5161 catgcat
//
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