名称: | pMG36e-P32-EGFP-nisI |
---|---|
别称: | |
基因长: | |
质粒宿主: | 乳酸菌 |
质粒用途: | 蛋白表达 |
片段类型: | ORF |
片段物种: | |
原核抗性: | Ery |
筛选标记: | Ery |
荧光标记: | |
启动子: | P32 |
复制子: | pWV01 |
拷贝数: | 低 |
感受态: | MC1061 |
温度: | 37度 |
背景载体: | |
正向引物: | |
反向引物: | |
诱导方式: |
1、本平台所有产品仅供科研使用,不能用于临床试验,包括人体口服、注射或者体外用药;
2、小包装质粒以模板价格出售,只保证基因部分测序正确,不保证质粒的表达等实验效果;
3、某些质粒即使是出品公司亦没有完整测过序,如关键部位的序列测序正确,亦认为是正确质粒;
4、小包装质粒产品的售后期为您收到产品之日起3个月,收到质粒后请及时转化操作,过期需重新购买;
5、质粒构建皆收取的是成本费用,所有质粒会分享给其他实验者,保密性质粒请提前告知并缴纳保密费用;
6、因科学研究为探索未知的活动,存在较大不确定性,故在任何情况下淼灵都不承担额外的连带责任;
7、发表[中文论文]请标注:产品名称(产品货号) 由武汉淼灵生物科技有限公司提供;
8、发表[英文论文]请标注:Product Name (Product Item Number) was obtained from MiaoLingBio,China.
操作说明
一、质粒干粉(负20度保存,保质期90天,常温运输)
1、收到干粉质粒后请先5000rpm离心1min,再加入20μl无菌水溶解质粒;
2、将相应的感受态于冰盒上解冻,取2μl质粒加至100μl感受态中,冰浴 30min;
3、42度热激60s,冰浴2min,加入900μl无抗LB培养基,180rpm震荡培养45min;
4、6000rpm离心3min,留100μl上清重悬,再涂至对应抗性的LB平板上;
5、倒置培养过夜后,如菌落过多则稀释质粒转化,如没有菌落则取10ul质粒转化;
6、挑单菌落至对应抗性的LB液体培养基中过夜震荡培养,根据需要提取质粒。
二、液体质粒(负20度保存,保质期90天,冰袋运输)
1、收到液体质粒后请甩动保存管,让液体质粒不要吸附在管壁上;
2、根据实验需求取适量质粒,其余保存于负20度冰箱。
三、甘油菌种(负80度保存,保质期90天,冰袋运输)
1、收到甘油菌种后请甩动保存管,让菌液不要吸附在管壁上;
2、用接种环在固体平板上四区划线,如有抗性就加对应抗性;
3、恒温培养箱倒置培养12~16h;
4、务必挑取单菌落接种于液体培养基中,如有抗性就加对应抗性;
5、摇床震荡培养过夜,根据实验需要培养菌种。
GGTGATTTCA
GAATCGAAAA
AAAGAGTTAT
GATTTCTCTG
ACAAAAGAGC
AAGATAAAAA
ATTAACAGAT
ATGGCGAAAC
AAAAAGGTTT
TTCAAAATCT
GCGGTTGCGG
CGTTAGCTAT
AGAAGAATAT
GCAAGAAAGG
AATCAGAACA
AAAAAAATAA
GCGAAAGCTC
GCGTTTTTAG
AAGGATACGA
GTTTTCGCTA
CTTGTTTTTG
ATAAGGTAAT
TATATCATGG
CTATTAAAAA
TACTAAAGCT
AGAAATTTTG
GATTTTTATT
ATATCCTGAC
TCAATTCCTA
ATGATTGGAA
AGAAAAATTA
GAGAGTTTGG
GCGTATCTAT
GGCTGTCAGT
CCTTTACACG
ATATGGACGA
AAAAAAAGAT
AAAGATACAT
GGAATAGTAG
TGATGTTATA
CGAAATGGAA
AGCACTATAA
AAAACCACAC
TATCACGTTA
TATATATTGC
ACGAAATCCT
GTAACAATAG
AAAGCGTTAG
GAACAAGATT
AAGCGAAAAT
TGGGGAATAG
TTCAGTTGCT
CATGTTGAGA
TACTTGATTA
TATCAAAGGT
TCATATGAAT
ATTTGACTCA
TGAATCAAAG
GACGCTATTG
CTAAGAATAA
ACATATATAC
GACAAAAAAG
ATATTTTGAA
CATTAATGAT
TTTGATATTG
ACCGCTATAT
AACACTTGAT
GAAAGCCAAA
AAAGAGAATT
GAAGAATTTA
CTTTTAGATA
TAGTGGATGA
CTATAATTTG
GTAAATACAA
AAGATTTAAT
GGCTTTTATT
CGCCTTAGGG
GAGCGGAGTT
TGGAATTTTA
AATACGAATG
ATGTAAAAGA
TATTGTTTCA
ACAAACTCTA
GCGCCTTTAG
ATTATGGTTT
GAGGGCAATT
ATCAGTGTGG
ATATAGAGCA
AGTTATGCAA
AGGTTCTTGA
TGCTGAAACG
GGGGAAATAA
AATGACAAAC
AAAGAAAAAG
AGTTATTTGC
TGAAAATGAG
GAATTAAAAA
AAGAAATTAA
GGACTTAAAA
GAGCGTATTG
AAAGATACAG
AGAAATGGAA
GTTGAATTAA
GTACAACAAT
AGATTTATTG
AGAGGAGGGA
TTATTGAATA
AATAAAAGCC
CCCTGACGAA
AGTCGAAGGG
GGTTTTTATT
TTGGTTTGAT
GTTGCGATTA
ATAGCAATAC
AATTGCAATA
AACAAAATGA
TCGACCTCGG
GACCCCTATC
TAGCGAACTT
TTAGAAAAGA
TATAAAACAT
CAGAGTATGG
ACAGTTGCGG
ATGTACTTCA
GAAAAGATTA
GATGTCTAAA
AAGCTAGCTT
TTTAGACATC
TAAATCTAGG
TACTAAAACA
ATTCATCCAG
TAAAATATAA
TATTTTATTT
TCTCCCAATC
AGGCTTGATC
CCCAGTAAGT
CAAAAAATAG
CTCGACATAC
TGTTCTTCCC
CGATCGACCC
GATTCACAAA
AAATAGGCAC
ACGAAAAACA
AGTTAAGGGA
TGCAGTTTAT
GCATCCCTTA
ACTTACTTAT
TAAATAATTT
ATAGCTATTG
AAAAGAGATA
AGAATTGTTC
AAAGCTAATA
TTGTTTAAAT
CGTCAATTCC
TGCATGTTTT
AAGGAATTGT
TAAATTGATT
TTTTGTAAAT
ATTTTCTTGT
ATTCTTTGTT
AACCCATTTC
ATAACGAAAT
AATTATACTT
TTGTTTATCT
TTGTGTGATA
TTCTTGATTT
TTTTCTACTT
AATCTGATAA
GTGAGCTATT
CACTTTAGGT
TTAGGATGAA
AATATTCTCT
TGGAACCATA
CTTAATATAG
AAATATCAAC
TTCTGCCATT
AAAAGTAATG
CCAATGAGCG
TTTTGTATTT
AATAATCTTT
TAGCAAACCC
GTATTCCACG
ATTAAATAAA
TCTCATTAGC
TATACTATCA
AAAACAATTT
TGCGTATTAT
ATCCGTACTT
ATGTTATAAG
GTATATTACC
ATATATTTTA
TAGGATTGGT
TTTTAGGAAA
TTTAAACTGC
AATATATCCT
TGTTTAAAAC
TTGGAAATTA
TCGTGATCAA
CAAGTTTATT
TTCTGTAGTT
TTGCATAATT
TATGGTCTAT
TTCAATGGCA
GTTACGAAAT
TACACCTCTT
TACTAATTCA
AGGGTAAAAT
GGCCTTTTCC
TGAGCCGATT
TCAAAGATAT
TATCATGTTC
ATTTAATCTT
ATATTTGTCA
TTATTTTATC
TATATTATGT
TTTGAAGTAA
TAAAGTTTTG
ACTGTGTTTT
ATATTTTTCT
CGTTCATTAT
AACCCTCTTT
AATTTGGTTA
TATGAATTTT
GCTTATTAAC
GATTCATTAT
AACCACTTAT
TTTTTGTTTG
GTTGATAATG
AACTGTGCTG
ATTACAAAAA
TACTAAAAAT
GCCCATATTT
TTTCCTCCTT
ATAAAATTAG
TATAATTATA
GCACGGTCGA
TCTTCTATAT
AAAAGATATA
TTATCTTATC
AGTATTGTCA
ATATATTCAA
GGCAATCTGC
CTCCTCATCC
TCTTCATCCT
CTTCGTCTTG
GTAGCTTTTT
AAATATGGGT
CGATCGAATT
CGGTCCTCGG
GATATGATAA
GATTAATAGT
TTTAGCTATT
AATCTTTTTT
TATTTTTATT
TAAGAATGGC
TTAATAAAGC
GGTTACTTTG
GATTTTTGTG
AGCTTGGACT
AGAAAAAAAC
TTCACAAAAT
GCTATACTAG
GTAGGTAAAA
AAATATTCGG
AGGAATTTTG
AAATGGCAAT
CGTTTCAGCA
GAAAAATTCG
TAATTCGAGC
TCATGGTGAG
CAAGGGCGAG
GAGCTGTTCA
CCGGGGTGGT
GCCCATCCTG
GTCGAGCTGG
ACGGCGACGT
AAACGGCCAC
AAGTTCAGCG
TGTCCGGCGA
GGGCGAGGGC
GATGCCACCT
ACGGCAAGCT
GACCCTGAAG
TTCATCTGCA
CCACCGGCAA
GCTGCCCGTG
CCCTGGCCCA
CCCTCGTGAC
CACCCTGACC
TACGGCGTGC
AGTGCTTCAG
CCGCTACCCC
GACCACATGA
AGCAGCACGA
CTTCTTCAAG
TCCGCCATGC
CCGAAGGCTA
CGTCCAGGAG
CGCACCATCT
TCTTCAAGGA
CGACGGCAAC
TACAAGACCC
GCGCCGAGGT
GAAGTTCGAG
GGCGACACCC
TGGTGAACCG
CATCGAGCTG
AAGGGCATCG
ACTTCAAGGA
GGACGGCAAC
ATCCTGGGGC
ACAAGCTGGA
GTACAACTAC
AACAGCCACA
ACGTCTATAT
CATGGCCGAC
AAGCAGAAGA
ACGGCATCAA
GGTGAACTTC
AAGATCCGCC
ACAACATCGA
GGACGGCAGC
GTGCAGCTCG
CCGACCACTA
CCAGCAGAAC
ACCCCCATCG
GCGACGGCCC
CGTGCTGCTG
CCCGACAACC
ACTACCTGAG
CACCCAGTCC
GCCCTGAGCA
AAGACCCCAA
CGAGAAGCGC
GATCACATGG
TCCTGCTGGA
GTTCGTGACC
GCCGCCGGGA
TCACTCTCGG
CATGGACGAG
CTGTACAAGT
CTGGTGGCGG
AGGCTCGGGC
GGAGGTGGGT
CGGGTGGCGG
CGGATCAATG
AGAAGATATT
TAATACTTAT
TGTGGCCTTA
ATAGGGATAA
CAGGTTTATC
AGGGTGTTAT
CAAACAAGTC
ATAAAAAGGT
GAGGTTTGAC
GAAGGAAGTT
ATACTAATTT
TATTTATGAT
AATAAATCGT
ATTTCGTAAC
TGATAAGGAG
ATTCCTCAGG
AGAACGTTAA
CAATTCCAAA
GTAAAATTTT
ATAAGCTGTT
GATTGTTGAC
ATGAAAAGTG
AGAAACTTTT
ATCAAGTAGC
AACAAAAATA
GTGTGACTTT
GGTCTTAAAT
AATATTTATG
AGGCTTCTGA
CAAGTCGCTA
TGTATGGGTA
TTAACGACAG
ATACTATAAG
ATACTTCCAG
AAAGTGATAA
GGGGGCGGTC
AAAGCTTTGA
GATTACAAAA
CTTTGATGTG
ACAAGCGATA
TTTCTGATGA
TAATTTTGTT
ATTGATAAAA
ATGATTCACG
AAAAATTGAC
TATATGGGAA
ATATTTACAG
TATATCGGAC
ACCACCGTAT
CTGATGAAGA
ATTGGGAGAA
TATCAGGATG
TTTTAGCTGA
AGTACGTGTG
TTTGATTCAG
TTAGTGGCAA
AAGTATCCCG
AGGTCTGAAT
GGGGGAGAAT
TGATAAGGAT
GGTTCAAATT
CCAAACAGAG
TAGGACGGAA
TGGGATTATG
GCGAAATCCA
TTCTATTAGA
GGAAAATCTC
TTACTGAAGC
ATTTGCCGTT
GAGATAAATG
ATGATTTTAA
GCTTGCAACG
AAGGTAGGAA
ACTAGCTGCA
GGCATGCAAG
CTTGCAAAGT
CTGAAAACGA
AGGTGGCAGC
TGCCGTTGAA
GCGGCCAAGA
CAGTTGGTAA
AGGCGACGGT
ACAACCGGTA
CTAGCGACAA
AGGCGGCGGT
CAAGGTACCC
CGGCGCTACG
ATATTTGGAG
TTGAGGTTCA
AAGTCAAATG
GTACTGATGA
CCGGTAAAAT
TTAATATTTT
GAACCTTGCT
TAGGCAGCTG
ACTTCACATT
GTTGAGATCA
GCTGCCTTTT
GCTTATAGTT
CATTGAGTAG
AAACGGTTCT
GTTGCGAAGT
TTGAAAATCA
AACGCAAGCT
CGATTTTTTA
TTAAAACGTC
TCAAAATCGT
TTCTGAGACG
TTTTAGCGTT
TATTTCGTTT
AGTTATCGGC
ATAATCGTTA
AAACAGGCGT
TATCGTAGCG
TAAAAGCCCT
TGAGCGTAGC
GTGGCTTTGC
AGCGAAGATG
TTGTCTGTTA
GATTATGAAA
GCCGATGACT
GAATGAAATA
ATAAGCGCAG
CGCCCTTCTA
TTTCGGTTGG
AGGAGGCTCA
AGGGAGTATG
AGGGAATGAA
ATTCCCTCAT
GGGTTTGATT
TTAAAAATTG
CTTGCAATTT
TGCCGAGCGG
TAGCGCTGGA
AAATTTTTGA
AAAAAATTTG
GAATTTGGAA
AAAAATGGGG
GGAAAGGAAG
CGAATTTTGC
TTCCGTACTA
CGACCCCCCA
TTAAGTGCCG
AGTGCCAATT
TTTGTGCCAA
AAACGCTCTA
TCCCAACTGG
CTCAAGGGTT
TAAGGGGTTT
TTCAATCGCC
AACGAATCGC
CAACGTTTTC
GCCAACGTTT
TTTATAAATC
TATATTTAAG
TAGCTTTATT
GTTGTTTTTA
TGATTACAAA
GTGATACACT
AACTTTATAA
AATTATTTGA
TTGGAGTTTT
TTAAT