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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20℃
- 保质期:
6个月
- 英文名:
pSV2-DHFR
- 库存:
大量
- 供应商:
联迈生物
- 规格:
1ug
质粒名称: pSV2-dhfr
其他名称:
质粒大小(bp): 4900
抗性: Ampicillin
质粒类型: 哺乳细胞载体;蛋白过表达质粒
是否病毒: Unspecified
启动子:SV 40
稳转或瞬转: Unspecified
组成型或诱导性: Unspecified
表达水平:
蛋白标签:C-DHFR ( mouse DHFR)
测序引物:
| 5’测序引物: | SV40pro-F: TATTTATGCAGAGGCCGAGG |
|---|---|
| 3’测序引物: | SV40pA-R: GAAATTTGTGATGCTATTGC |
GenBank号:
其他属性:
pSV2-DHFR载体含有的是小鼠的DHFR基因,同时该载体是哺乳动物细胞和大肠杆菌穿梭质粒,在DHFR细胞系中,可以通过甲氨蝶呤对转染的细胞进行筛选。
质粒图谱:
质粒序列:
| 1 | cTGTGGAATG | TGTGTCAGTT | AGGGTGTGGA | AAGTCCCCAG | GCTCCCCAGC | 50 |
| 51 | AGGCAGAAGT | ATGCAAAGCA | TGCATCTCAA | TTAGTCAGCA | ACCAGGTGTG | 100 |
| 101 | GAAAGTCCCC | AGGCTCCCCA | GCAGGCAGAA | GTATGCAAAG | CATGCATCTC | 150 |
| 151 | AATTAGTCAG | CAACCATAGT | CCCGCCCCTA | ACTCCGCCCA | TCCCGCCCCT | 200 |
| 201 | AACTCCGCCC | AGTTCCGCCC | ATTCTCCGCC | CCATGGCTGA | CTAATTTTTT | 250 |
| 251 | TTATTTATGC | AGAGGCCGAG | GCCGCCTCGG | CCTCTGAGCT | ATTCCAGAAG | 300 |
| 301 | TAGTGAGGAG | GCTTTTTTGG | AGGCCTAGGC | TTTTGCAAAA | AGCTTTATCC | 350 |
| 351 | CCGCTGCCAT | CATGGTTCGA | CCATTGAACT | GCATCGTCGC | CGTGTCCCAA | 400 |
| 401 | GATATGGGGA | TTGGCAAGAA | CGGAGACCTA | CCCTGGCCTC | CGCTCAGGAA | 450 |
| 451 | CGAGTTCAAG | TACTTCCAAA | GAATGACCAC | AACCTCTTCA | GTGGAAGGTA | 500 |
| 501 | AACAGAATCT | GGTGATTATG | GGTAGGAAAA | CCTGGTTCTC | CATTCCTGAG | 550 |
| 551 | AAGAATCGAC | CTTTAAAGGA | CAGAATTAAT | ATAGTTCTCA | GTAGAGAACT | 600 |
| 601 | CAAAGAACCA | CCACGAGGAG | CTCATTTTCT | TGCCAAAAGT | TTGGATGATG | 650 |
| 651 | CCTTAAGACT | TATTGAACAA | CCGGAATTGG | CAAGTAAAGT | AGACATGGTT | 700 |
| 701 | TGGATAGTCG | GAGGCAGTTC | TGTTTACCAG | GAAGCCATGA | ATCAACCAGG | 750 |
| 751 | CCACCTCAGA | CTCTTTGTGA | CAAGGATCAT | GCAGGAATTT | GAAAGTGACA | 800 |
| 801 | CGTTTTTCCC | AGAAATTGAT | TTGGGGAAAT | ATAAACTTCT | CCCAGAATAC | 850 |
| 851 | CCAGGCGTCC | TCTCTGAGGT | CCAGGAGGAA | AAAGGCATCA | AGTATAAGTT | 900 |
| 901 | TGAAGTCTAC | GAGAAGAAAG | ACTAACAGGA | AGATGCTTTC | AAGTTCTCTG | 950 |
| 951 | CTCCCCTCCT | AAAGCTATGC | ATTTTTATAA | GACCATGGGA | CTTTTGCTGG | 1000 |
| 1001 | CTTTAGATCT | TTGTGAAGGA | ACCTTACTTC | TGTGGTGTGA | CATAATTGGA | 1050 |
| 1051 | CAAACTACCT | ACAGAGATTT | AAAGCTCTAA | GGTAAATATA | AAATTTTTAA | 1100 |
| 1101 | GTGTATAATG | TGTTAAACTA | CTGATTCTAA | TTGTTTGTGT | ATTTTAGATT | 1150 |
| 1151 | CCAACCTATG | GAACTGATGA | ATGGGAGCAG | TGGTGGAATG | CCTTTAATGA | 1200 |
| 1201 | GGAAAACCTG | TTTTGCTCAG | AAGAAATGCC | ATCTAGTGAT | GATGAGGCTA | 1250 |
| 1251 | CTGCTGACTC | TCAACATTCT | ACTCCTCCAA | AAAAGAAGAG | AAAGGTAGAA | 1300 |
| 1301 | GACCCCAAGG | ACTTTCCTTC | AGAATTGCTA | AGTTTTTTGA | GTCATGCTGT | 1350 |
| 1351 | GTTTAGTAAT | AGAACTCTTG | CTTGCTTTGC | TATTTACACC | ACAAAGGAAA | 1400 |
| 1401 | AAGCTGCACT | GCTATACAAG | AAAATTATGG | AAAAATATTC | TGTAACCTTT | 1450 |
| 1451 | ATAAGTAGGC | ATAACAGTTA | TAATCATAAC | ATACTGTTTT | TTCTTACTCC | 1500 |
| 1501 | ACACAGGCAT | AGAGTGTCTG | CTATTAATAA | CTATGCTCAA | AAATTGTGTA | 1550 |
| 1551 | CCTTTAGCTT | TTTAATTTGT | AAAGGGGTTA | ATAAGGAATA | TTTGATGTAT | 1600 |
| 1601 | AGTGCCTTGA | CTAGAGATCA | TAATCAGCCA | TACCACATTT | GTAGAGGTTT | 1650 |
| 1651 | TACTTGCTTT | AAAAAACCTC | CCACACCTCC | CCCTGAACCT | GAAACATAAA | 1700 |
| 1701 | ATGAATGCAA | TTGTTGTTGT | TAACTTGTTT | ATTGCAGCTT | ATAATGGTTA | 1750 |
| 1751 | CAAATAAAGC | AATAGCATCA | CAAATTTCAC | AAATAAAGCA | TTTTTTTCAC | 1800 |
| 1801 | TGCATTCTAG | TTGTGGTTTG | TCCAAACTCA | TCAATGTATC | TTATCATGTC | 1850 |
| 1851 | TGGATCCCCA | GGAAGCTCCT | CTGTGTCCTC | ATAAACCCTA | ACCTCCTCTA | 1900 |
| 1901 | CTTGAGAGGA | CATTCCAATC | ATAGGCTGCC | CATCCACCCT | CTGTGTCCTC | 1950 |
| 1951 | CTGTTAATTA | GGTCACTTAA | CAAAAAGGAA | ATTGGGTAGG | GGTTTTTCAC | 2000 |
| 2001 | AGACCGCTTT | CTAAGGGTAA | TTTTAAAATA | TCTGGGAAGT | CCCTTCCACT | 2050 |
| 2051 | GCTGTGTTCC | AGAAGTGTTG | GTAAACAGCC | CACAAATGTC | AACAGCAGAA | 2100 |
| 2101 | ACATACAAGC | TGTCAGCTTT | GCACAAGGGC | CCAACACCCT | GCTCATCAAG | 2150 |
| 2151 | AAGCACTGTG | GTTGCTGTGT | TAGTAATGTG | CAAAACAGGA | GGCACATTTT | 2200 |
| 2201 | CCCCACCTGT | GTAGGTTCCA | AAATATCTAG | TGTTTTCATT | TTTACTTGGA | 2250 |
| 2251 | TCAGGAACCC | AGCACTCCAC | TGGATAAGCA | TTATCCTTAT | CCAAAACAGC | 2300 |
| 2301 | CTTGTGGTCA | GTGTTCATCT | GCTGACTGTC | AACTGTAGCA | TTTTTTGGGG | 2350 |
| 2351 | TTACAGTTTG | AGCAGGATAT | TTGGTCCTGT | AGTTTGCTAA | CACACCCTGC | 2400 |
| 2401 | AGCTCCAAAG | GTTCCCCACC | AACAGCAAAA | AAATGAAAAT | TTGACCCTTG | 2450 |
| 2451 | AATGGGTTTT | CCAGCACCAT | TTTCATGAGT | TTTTTGTGTC | CCTGAATGCA | 2500 |
| 2501 | AGTTTAACAT | AGCAGTTACC | CCAATAACCT | CAGTTTTAAC | AGTAACAGCT | 2550 |
| 2551 | TCCCACATCA | AAATATTTCC | ACAGGTTAAG | TCCTCATTTA | AATTAGGCAA | 2600 |
| 2601 | AGGAATTCTT | GAAGACGAAA | GGGCCTCGTG | ATACGCCTAT | TTTTATAGGT | 2650 |
| 2651 | TAATGTCATG | ATAATAATGG | TTTCTTAGAC | GTCAGGTGGC | ACTTTTCGGG | 2700 |
| 2701 | GAAATGTGCG | CGGAACCCCT | ATTTGTTTAT | TTTTCTAAAT | ACATTCAAAT | 2750 |
| 2751 | ATGTATCCGC | TCATGAGACA | ATAACCCTGA | TAAATGCTTC | AATAATATTG | 2800 |
| 2801 | AAAAAGGAAG | AGTATGAGTA | TTCAACATTT | CCGTGTCGCC | CTTATTCCCT | 2850 |
| 2851 | TTTTTGCGGC | ATTTTGCCTT | CCTGTTTTTG | CTCACCCAGA | AACGCTGGTG | 2900 |
| 2901 | AAAGTAAAAG | ATGCTGAAGA | TCAGTTGGGT | GCACGAGTGG | GTTACATCGA | 2950 |
| 2951 | ACTGGATCTC | AACAGCGGTA | AGATCCTTGA | GAGTTTTCGC | CCCGAAGAAC | 3000 |
| 3001 | GTTTTCCAAT | GATGAGCACT | TTTAAAGTTC | TGCTATGTGG | CGCGGTATTA | 3050 |
| 3051 | TCCCGTGTTG | ACGCCGGGCA | AGAGCAACTC | GGTCGCCGCA | TACACTATTC | 3100 |
| 3101 | TCAGAATGAC | TTGGTTGAGT | ACTCACCAGT | CACAGAAAAG | CATCTTACGG | 3150 |
| 3151 | ATGGCATGAC | AGTAAGAGAA | TTATGCAGTG | CTGCCATAAC | CATGAGTGAT | 3200 |
| 3201 | AACACTGCGG | CCAACTTACT | TCTGACAACG | ATCGGAGGAC | CGAAGGAGCT | 3250 |
| 3251 | AACCGCTTTT | TTGCACAACA | TGGGGGATCA | TGTAACTCGC | CTTGATCGTT | 3300 |
| 3301 | GGGAACCGGA | GCTGAATGAA | GCCATACCAA | ACGACGAGCG | TGACACCACG | 3350 |
| 3351 | ATGCCTGCAG | CAATGGCAAC | AACGTTGCGC | AAACTATTAA | CTGGCGAACT | 3400 |
| 3401 | ACTTACTCTA | GCTTCCCGGC | AACAATTAAT | AGACTGGATG | GAGGCGGATA | 3450 |
| 3451 | AAGTTGCAGG | ACCACTTCTG | CGCTCGGCCC | TTCCGGCTGG | CTGGTTTATT | 3500 |
| 3501 | GCTGATAAAT | CTGGAGCCGG | TGAGCGTGGG | TCTCGCGGTA | TCATTGCAGC | 3550 |
| 3551 | ACTGGGGCCA | GATGGTAAGC | CCTCCCGTAT | CGTAGTTATC | TACACGACGG | 3600 |
| 3601 | GGAGTCAGGC | AACTATGGAT | GAACGAAATA | GACAGATCGC | TGAGATAGGT | 3650 |
| 3651 | GCCTCACTGA | TTAAGCATTG | GTAACTGTCA | GACCAAGTTT | ACTCATATAT | 3700 |
| 3701 | ACTTTAGATT | GATTTAAAAC | TTCATTTTTA | ATTTAAAAGG | ATCTAGGTGA | 3750 |
| 3751 | AGATCCTTTT | TGATAATCTC | ATGACCAAAA | TCCCTTAACG | TGAGTTTTCG | 3800 |
| 3801 | TTCCACTGAG | CGTCAGACCC | CGTAGAAAAG | ATCAAAGGAT | CTTCTTGAGA | 3850 |
| 3851 | TCCTTTTTTT | CTGCGCGTAA | TCTGCTGCTT | GCAAACAAAA | AAACCACCGC | 3900 |
| 3901 | TACCAGCGGT | GGTTTGTTTG | CCGGATCAAG | AGCTACCAAC | TCTTTTTCCG | 3950 |
| 3951 | AAGGTAACTG | GCTTCAGCAG | AGCGCAGATA | CCAAATACTG | TCCTTCTAGT | 4000 |
| 4001 | GTAGCCGTAG | TTAGGCCACC | ACTTCAAGAA | CTCTGTAGCA | CCGCCTACAT | 4050 |
| 4051 | ACCTCGCTCT | GCTAATCCTG | TTACCAGTGG | CTGCTGCCAG | TGGCGATAAG | 4100 |
| 4101 | TCGTGTCTTA | CCGGGTTGGA | CTCAAGACGA | TAGTTACCGG | ATAAGGCGCA | 4150 |
| 4151 | GCGGTCGGGC | TGAACGGGGG | GTTCGTGCAC | ACAGCCCAGC | TTGGAGCGAA | 4200 |
| 4201 | CGACCTACAC | CGAACTGAGA | TACCTACAGC | GTGAGCTATG | AGAAAGCGCC | 4250 |
| 4251 | ACGCTTCCCG | AAGGGAGAAA | GGCGGACAGG | TATCCGGTAA | GCGGCAGGGT | 4300 |
| 4301 | CGGAACAGGA | GAGCGCACGA | GGGAGCTTCC | AGGGGGAAAC | GCCTGGTATC | 4350 |
| 4351 | TTTATAGTCC | TGTCGGGTTT | CGCCACCTCT | GACTTGAGCG | TCGATTTTTG | 4400 |
| 4401 | TGATGCTCGT | CAGGGGGGCG | GAGCCTATGG | AAAAACGCCA | GCAACGCGGC | 4450 |
| 4451 | CTTTTTACGG | TTCCTGGCCT | TTTGCTGGCC | TTTTGCTCAC | ATGTTCTTTC | 4500 |
| 4501 | CTGCGTTATC | CCCTGATTCT | GTGGATAACC | GTATTACCGC | CTTTGAGTGA | 4550 |
| 4551 | GCTGATACCG | CTCGCCGCAG | CCGAACGACC | GAGCGCAGCG | AGTCAGTGAG | 4600 |
| 4601 | CGAGGAAGCG | GAAGAGCGCC | TGATGCGGTA | TTTTCTCCTT | ACGCATCTGT | 4650 |
| 4651 | GCGGTATTTC | ACACCGCATA | TGGTGCACTC | TCAGTACAAT | CTGCTCTGAT | 4700 |
| 4701 | GCCGCATAGT | TAAGCCAGTA | TACACTCCGC | TATCGCTACG | TGACTGGGTC | 4750 |
| 4751 | ATGGCTGCGC | CCCGACACCC | GCCAACACCC | GCTGACGCGC | CCTGACGGGC | 4800 |
| 4801 | TTGTCTGCTC | CCGGCATCCG | CTTACAGACA | AGCTGTGACC | GTCTCCGGGA | 4850 |
| 4851 | GCTGCATGTG | TCAGAGGTTT | TCACCGTCAT | CACCGAAACG | CGCGAGGCAG | 4900 |
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文献和实验Using the DHFR Heat-Inducible Degron for Protein Inactivation in Schizosaccharomyces pombe
Inactivating a specific protein in vivo can yield important information about its function. One strategy previously developed inSaccharomyces cerevisiae by the Varshavsky group involves fusing a degron, derived from mouse dihydrofolate
细胞系等,该细胞有一定自发转化倾向,一般在转染前一天接种细胞,接种密度为2×104 /cm2 ,用含10%胎牛血清的DMEM液,37℃、5%CO2 培养,待细胞占50~70%瓶底面积时,用于转染试验。 (3)DNA-磷酸钙沉淀物的制备 ①将供体细胞DNA和PSV2-neo质粒载体DNA用TE配制成40mg/L的DNA溶液,同时向供体细胞DNA液200μl中加入带基因neo质粒DNA液(20mg/L)220μl和2×HBS 250μl(PSV2-neo为1~2mg/L的使用量)。 ②取500μl上述
Cytogenetic Characterization of Recombinant Cells
; therefore a selectable system is desirable to discriminate cells that have integrated the foreign DNA. Chinese hamster ovary (CHO) cells are most commonly used because of the availability of both a dihydrofolate reductase (DHFR)-deficient mutant cell line (CHO-DUKX
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