- 详细信息
- 文献和实验
- 技术资料
- 库存:
大量
- 英文名:
pBR322
- 保质期:
1年
- 供应商:
再康生物
- 保存条件:
低温运输,-20℃保存
质粒编号: 60908-1110y
质粒名称: pBR322
其他名称: Unknown
质粒大小(bp): 4361
抗性: Ampicillin
质粒类型: 克隆载体
是否病毒: Unspecified
启动子: Unspecified
稳转或瞬转: Unspecified
组成型或诱导性: Unspecified
表达水平: Unspecified
蛋白标签: Unspecified
测序引物:
GenBank号: M10785, M10786, M33694
其他属性:
Size is 4363 for Boehringer, BRL, Promega, Sigma. Size is 4362 for USB. Promoter P1 is artificially created by the ligation of two different DNA fragments to create pBR322. Promoter P2 in the same region as P1, but it is on the opposite strand and initiates transcription in the direction of the tetracycline resistance gene. Promoter P3 is the natural promoter for the beta-lactamase gene. Medium is 1227 LB plus ampicillin. The circular sequence is numbered such that 0 is the middle of the unique EcoRI site and the count increases first through the tet genes, the pMB1 material, and finally through the Tn3 region. NCBI gi: 58257 Hosts: E.coli HB101, E.coli C600, E.coli RR1, E.coli. Related vectors: pSC101. Also, from Bolivar et al., Gene, 77: pBR322 is derived from pBR313, is a relaxed replicating plasmid, does not produce and is sensitive to colicin E1, and carries resistance genes to the antibiotics ampicillin (Ap) and tetracycline (Tc). The antibiotic-resistant genes on pBR322 are not transposable. The vector pBR322 was constructed in order to have a plasmid with a single PstI site, located in the ampicillin-resistant gene (Apr), in addition to four unique restriction sites, EcoRI, HindIII, BamHI and SalI.
质粒图谱:
质粒序列:
| 1 | ttctcatgtt | tgacagctta | tcatcgataa | gctttaatgc | ggtagtttat | 50 |
| 51 | cacagttaaa | ttgctaacgc | agtcaggcac | cgtgtatgaa | atctaacaat | 100 |
| 101 | gcgctcatcg | tcatcctcgg | caccgtcacc | ctggatgctg | taggcatagg | 150 |
| 151 | cttggttatg | ccggtactgc | cgggcctctt | gcgggatatc | gtccattccg | 200 |
| 201 | acagcatcgc | cagtcactat | ggcgtgctgc | tagcgctata | tgcgttgatg | 250 |
| 251 | caatttctat | gcgcacccgt | tctcggagca | ctgtccgacc | gctttggccg | 300 |
| 301 | ccgcccagtc | ctgctcgctt | cgctacttgg | agccactatc | gactacgcga | 350 |
| 351 | tcatggcgac | cacacccgtc | ctgtggatcc | tctacgccgg | acgcatcgtg | 400 |
| 401 | gccggcatca | ccggcgccac | aggtgcggtt | gctggcgcct | atatcgccga | 450 |
| 451 | catcaccgat | ggggaagatc | gggctcgcca | cttcgggctc | atgagcgctt | 500 |
| 501 | gtttcggcgt | gggtatggtg | gcaggccccg | tggccggggg | actgttgggc | 550 |
| 551 | gccatctcct | tgcatgcacc | attccttgcg | gcggcggtgc | tcaacggcct | 600 |
| 601 | caacctacta | ctgggctgct | tcctaatgca | ggagtcgcat | aagggagagc | 650 |
| 651 | gtcgaccgat | gcccttgaga | gccttcaacc | cagtcagctc | cttccggtgg | 700 |
| 701 | gcgcggggca | tgactatcgt | cgccgcactt | atgactgtct | tctttatcat | 750 |
| 751 | gcaactcgta | ggacaggtgc | cggcagcgct | ctgggtcatt | ttcggcgagg | 800 |
| 801 | accgctttcg | ctggagcgcg | acgatgatcg | gcctgtcgct | tgcggtattc | 850 |
| 851 | ggaatcttgc | acgccctcgc | tcaagccttc | gtcactggtc | ccgccaccaa | 900 |
| 901 | acgtttcggc | gagaagcagg | ccattatcgc | cggcatggcg | gccgacgcgc | 950 |
| 951 | tgggctacgt | cttgctggcg | ttcgcgacgc | gaggctggat | ggccttcccc | 1000 |
| 1001 | attatgattc | ttctcgcttc | cggcggcatc | gggatgcccg | cgttgcaggc | 1050 |
| 1051 | catgctgtcc | aggcaggtag | atgacgacca | tcagggacag | cttcaaggat | 1100 |
| 1101 | cgctcgcggc | tcttaccagc | ctaacttcga | tcactggacc | gctgatcgtc | 1150 |
| 1151 | acggcgattt | atgccgcctc | ggcgagcaca | tggaacgggt | tggcatggat | 1200 |
| 1201 | tgtaggcgcc | gccctatacc | ttgtctgcct | ccccgcgttg | cgtcgcggtg | 1250 |
| 1251 | catggagccg | ggccacctcg | acctgaatgg | aagccggcgg | cacctcgcta | 1300 |
| 1301 | acggattcac | cactccaaga | attggagcca | atcaattctt | gcggagaact | 1350 |
| 1351 | gtgaatgcgc | aaaccaaccc | ttggcagaac | atatccatcg | cgtccgccat | 1400 |
| 1401 | ctccagcagc | cgcacgcggc | gcatctcggg | cagcgttggg | tcctggccac | 1450 |
| 1451 | gggtgcgcat | gatcgtgctc | ctgtcgttga | ggacccggct | aggctggcgg | 1500 |
| 1501 | ggttgcctta | ctggttagca | gaatgaatca | ccgatacgcg | agcgaacgtg | 1550 |
| 1551 | aagcgactgc | tgctgcaaaa | cgtctgcgac | ctgagcaaca | acatgaatgg | 1600 |
| 1601 | tcttcggttt | ccgtgtttcg | taaagtctgg | aaacgcggaa | gtcagcgccc | 1650 |
| 1651 | tgcaccatta | tgttccggat | ctgcatcgca | ggatgctgct | ggctaccctg | 1700 |
| 1701 | tggaacacct | acatctgtat | taacgaagcg | ctggcattga | ccctgagtga | 1750 |
| 1751 | tttttctctg | gtcccgccgc | atccataccg | ccagttgttt | accctcacaa | 1800 |
| 1801 | cgttccagta | accgggcatg | ttcatcatca | gtaacccgta | tcgtgagcat | 1850 |
| 1851 | cctctctcgt | ttcatcggta | tcattacccc | catgaacaga | aatccccctt | 1900 |
| 1901 | acacggaggc | atcagtgacc | aaacaggaaa | aaaccgccct | taacatggcc | 1950 |
| 1951 | cgctttatca | gaagccagac | attaacgctt | ctggagaaac | tcaacgagct | 2000 |
| 2001 | ggacgcggat | gaacaggcag | acatctgtga | atcgcttcac | gaccacgctg | 2050 |
| 2051 | atgagcttta | ccgcagctgc | ctcgcgcgtt | tcggtgatga | cggtgaaaac | 2100 |
| 2101 | ctctgacaca | tgcagctccc | ggagacggtc | acagcttgtc | tgtaagcgga | 2150 |
| 2151 | tgccgggagc | agacaagccc | gtcagggcgc | gtcagcgggt | gttggcgggt | 2200 |
| 2201 | gtcggggcgc | agccatgacc | cagtcacgta | gcgatagcgg | agtgtatact | 2250 |
| 2251 | ggcttaacta | tgcggcatca | gagcagattg | tactgagagt | gcaccatatg | 2300 |
| 2301 | cggtgtgaaa | taccgcacag | atgcgtaagg | agaaaatacc | gcatcaggcg | 2350 |
| 2351 | ctcttccgct | tcctcgctca | ctgactcgct | gcgctcggtc | gttcggctgc | 2400 |
| 2401 | ggcgagcggt | atcagctcac | tcaaaggcgg | taatacggtt | atccacagaa | 2450 |
| 2451 | tcaggggata | acgcaggaaa | gaacatgtga | gcaaaaggcc | agcaaaaggc | 2500 |
| 2501 | caggaaccgt | aaaaaggccg | cgttgctggc | gtttttccat | aggctccgcc | 2550 |
| 2551 | cccctgacga | gcatcacaaa | aatcgacgct | caagtcagag | gtggcgaaac | 2600 |
| 2601 | ccgacaggac | tataaagata | ccaggcgttt | ccccctggaa | gctccctcgt | 2650 |
| 2651 | gcgctctcct | gttccgaccc | tgccgcttac | cggatacctg | tccgcctttc | 2700 |
| 2701 | tcccttcggg | aagcgtggcg | ctttctcata | gctcacgctg | taggtatctc | 2750 |
| 2751 | agttcggtgt | aggtcgttcg | ctccaagctg | ggctgtgtgc | acgaaccccc | 2800 |
| 2801 | cgttcagccc | gaccgctgcg | ccttatccgg | taactatcgt | cttgagtcca | 2850 |
| 2851 | acccggtaag | acacgactta | tcgccactgg | cagcagccac | tggtaacagg | 2900 |
| 2901 | attagcagag | cgaggtatgt | aggcggtgct | acagagttct | tgaagtggtg | 2950 |
| 2951 | gcctaactac | ggctacacta | gaaggacagt | atttggtatc | tgcgctctgc | 3000 |
| 3001 | tgaagccagt | taccttcgga | aaaagagttg | gtagctcttg | atccggcaaa | 3050 |
| 3051 | caaaccaccg | ctggtagcgg | tggttttttt | gtttgcaagc | agcagattac | 3100 |
| 3101 | gcgcagaaaa | aaaggatctc | aagaagatcc | tttgatcttt | tctacggggt | 3150 |
| 3151 | ctgacgctca | gtggaacgaa | aactcacgtt | aagggatttt | ggtcatgaga | 3200 |
| 3201 | ttatcaaaaa | ggatcttcac | ctagatcctt | ttaaattaaa | aatgaagttt | 3250 |
| 3251 | taaatcaatc | taaagtatat | atgagtaaac | ttggtctgac | agttaccaat | 3300 |
| 3301 | gcttaatcag | tgaggcacct | atctcagcga | tctgtctatt | tcgttcatcc | 3350 |
| 3351 | atagttgcct | gactccccgt | cgtgtagata | actacgatac | gggagggctt | 3400 |
| 3401 | accatctggc | cccagtgctg | caatgatacc | gcgagaccca | cgctcaccgg | 3450 |
| 3451 | ctccagattt | atcagcaata | aaccagccag | ccggaagggc | cgagcgcaga | 3500 |
| 3501 | agtggtcctg | caactttatc | cgcctccatc | cagtctatta | attgttgccg | 3550 |
| 3551 | ggaagctaga | gtaagtagtt | cgccagttaa | tagtttgcgc | aacgttgttg | 3600 |
| 3601 | ccattgctgc | aggcatcgtg | gtgtcacgct | cgtcgtttgg | tatggcttca | 3650 |
| 3651 | ttcagctccg | gttcccaacg | atcaaggcga | gttacatgat | cccccatgtt | 3700 |
| 3701 | gtgcaaaaaa | gcggttagct | ccttcggtcc | tccgatcgtt | gtcagaagta | 3750 |
| 3751 | agttggccgc | agtgttatca | ctcatggtta | tggcagcact | gcataattct | 3800 |
| 3801 | cttactgtca | tgccatccgt | aagatgcttt | tctgtgactg | gtgagtactc | 3850 |
| 3851 | aaccaagtca | ttctgagaat | agtgtatgcg | gcgaccgagt | tgctcttgcc | 3900 |
| 3901 | cggcgtcaac | acgggataat | accgcgccac | atagcagaac | tttaaaagtg | 3950 |
| 3951 | ctcatcattg | gaaaacgttc | ttcggggcga | aaactctcaa | ggatcttacc | 4000 |
| 4001 | gctgttgaga | tccagttcga | tgtaacccac | tcgtgcaccc | aactgatctt | 4050 |
| 4051 | cagcatcttt | tactttcacc | agcgtttctg | ggtgagcaaa | aacaggaagg | 4100 |
| 4101 | caaaatgccg | caaaaaaggg | aataagggcg | acacggaaat | gttgaatact | 4150 |
| 4151 | catactcttc | ctttttcaat | attattgaag | catttatcag | ggttattgtc | 4200 |
| 4201 | tcatgagcgg | atacatattt | gaatgtattt | agaaaaataa | acaaataggg | 4250 |
| 4251 | gttccgcgca | catttccccg | aaaagtgcca | cctgacgtct | aagaaaccat | 4300 |
| 4301 | tattatcatg | acattaacct | ataaaaatag | gcgtatcacg | aggccctttc | 4350 |
| 4351 | gtcttcaaga | a | 4361 |
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文献和实验Back to Basics: pBR322 and Protein Expression Systems in E. coli
plasmid-borne elements and their interaction with the particular host strain will influence the overall expression system and final productivity (3 ) (see Chapters 1 –3 ). Plasmid vector pBR322 (4 ) is a well-established multipurpose cloning
1),尿嘧啶合成酶基因(URA3),亮氨酸合成酶基因(LEU2),组氨酸合成酶基因(HIS4)等。 3) 生化标记基因。这类标记基因的表达产物可催化某些易检测的生化反应,常用的基因有β-半乳苷酶基因(lacZ)、葡萄糖苷酸酶基因(GUS)、氯霉素乙酰转移酶基因(CAT)。 为了进一步说明载体的结构和各个结构的功能,下面举一个在基因工程中使用比较频繁的大肠杆菌载体—pBR322作为例子。 大肠杆菌载体pBR322 载体pBR322是由Bolivar 等人于1977年构建的。该载
都中止。但松弛型质粒仍继续进行复制。因此在增殖松弛型质粒时,总是要让宿主菌经氯霉素处理(在培养液中加入适量的氯霉素)抑制蛋白质的合成。图23-3 pBR322的基本结构(一)大肠杆菌质粒应用的最广泛的质粒载体是pBR322,它属松弛型质粒,有抗氨苄青霉素和抗四环素两个抗性基因可作为标记基因,有许多种常用的限制酶的切点。它全长4352个核苷酸,排列顺序已全部测定,基本结构如图23-3。当需要用pBR322来克隆BamHI酶切的一个DNA片段时,一般的做法是如图23-4所示。图23-4 pBR322克隆
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