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1、常见限制性内切酶识别序列（酶切位点）（BamHI、EcoRI、HindIII、NdeI、XhoI 等）Time：2009-10-22 PM 15:38 Author:bioer Hits: 7681 times在分子克隆实验中，限制性内切酶是必不可少的工具酶。无论是构建克隆载体还是表达载体，要根据载体选择合适的内切酶（当然，使用T 载就不必考虑了）。先将引物设计好，然后添加酶切识别序列到引物5 端。常用的内切酶比 如 BamHI、 EcoRI、 HindIII 、 NdeI、XhoI 等可能你都已经记住了它们的识别序列，不过 为了保险起见，还是得查证一下。下面是一些常用的II型内切酶的识别序列
2、，仅供参考。先介绍一下什么是II型内 切酶吧。The Type II restriction systems typically contain individual restriction enzymes and modification enzymes encoded by separate genes. The Type II restriction enzymes typically recognize specific DNA sequences and cleave at constant positions at or close to that sequence to prod
3、uce 5-phosphates and 3-hydroxyls. Usually they require Mg 2+ ions as a cofactor, although some have more exotic requirements. The methyltransferases usually recognize the same sequence although some are more promiscuous. Three types of DNA methyltransferases have been found as part of Type II R-M sy
4、stems forming either C5-methylcytosine, N4-methylcytosine or N6-methyladenine.酶类型识别序列TypeIIrestrictionApaIenzyme5GGGC3C 3TypeIIrestriction5 GAGATCC 3BamHIenzymeTypeIIrestrictionBglIIenzyme5 AAGATCT 3TypeIIrestrictionEcoRIenzyme5 GAAATTC 3TypeIIrestrictionHindIIIenzyme5 AAAGCTT 3TypeIIrestrictionKpnI
5、enzyme5 GGTA3C 3NcoITypeenzymeIIrestriction5 CACATGG 3NdeITypeenzymeIIrestriction5 CAATATG 3NheITypeenzymeIIrestriction5 GACTAGC 3NotITypeenzymeIIrestriction5 GCAGGCCGC 3SacITypeenzymeIIrestriction5 GAGCTAC 3SalITypeenzymeIIrestriction5 GATCGAC 3SphITypeenzymeIIrestriction5 GCATGAC 3XbaITypeenzymeII
6、restriction5 TACTAGA 3XhoITypeenzymeIIrestriction5 CATCGAG 3要查找更多内切酶的识别序列，你还可以选择下面几种方法1. 查你所使用的内切酶的公司的目录或者网站；2. 用软件如：Primer Premier5.o或Bioedit等，这些软件均提供了内切酶识别序列的信息;3. 推荐到 NEB 的 REBASE 数据库去查（网址 http:/ SequenceChain LengthQ/o Cleavage2 hr20 hrAccIGGTCGACC800CGGTCGACCG1000CCGGTCGACCGG1200AflUICACATGTG80
7、0CCACATGTGG109090CCCACATGTGGG129090AsciGGCGCGCC89090AGGCGCGCCT109090TTGGCGCGCCAA129090AvalCCCCGGGG85090CCCCCGGGGG109090TCCCCCGGGGGA129090BamHICGGATCCG81025CGGGATCCCG109090CGCGGATCCGCG129090BginCAGATCTG800gaagatcttc107590ggaagatcttcc122590BSSHIIGGCGCGCC800AGGCGCGCCT1000TTGGCG CGCCAA125090BstEIiGGGT(A
8、/T)ACCC9010BstXIAACTG CAGAACCAATGCATT G G2200AAAACT G CAGCCAAT GCATTG G 4A242550CT GCAGAACCAAT GC ATT G G AT GCAT272590ClalCATCGATG800GATCGATC800CCATCGATGG109090CCCATCGATGGG125050EcoRIGGAATTCC89090CGGAATTCCG109090CCGGAATTCCGG129090Haelll HindlllKpnlMlulNcolNdelNhelNoUNsilPacIPmelGGGGCCCCAGCGGCCGCTTT
9、GCGGCCGCAACAAGCTTGCCAAGCTTGGCCCAAGCTTGGGGGGTACCCGGGGTACCCCCGGGGTACCCCGGACGCGTCCGACGCGTCGCCCATGGGCATGCCATGGCATGCCATATGGCCCATATGGGCGCCATATGGCGGGGTTTCATAT G AAACCCGGAATTCCATAT G GAATTCCGGGAATTCCATATGGAATTCCCGGCTAGCCCGGCTAGCCGCTAGCTAGCTAG890901090901290908001000121075800109090129090800102550800145075800
10、100012001800207590227590800101025121050bCGGLCbLAA1200ATTTGCGGCCGCTTTA161010AAATATGCGGCCG CTATAAA201010ataagaatgcggccgctaaactat242590aaggaaaaaagcggccgcaaaaggaaaa282590TGCATGCATGCA121090CCAAT GCATT G GTT CT G CAGTT229090ttaattaa800gttaattaac10025CCTTAATTAAGG12090gtttaaac800GGTTTAAACC10025GGGTTTAAACCC1
11、2050AGC1 1 1G1 1 1AAAC GGCG CGCCGG247590PstrGCTG8TGCACTGCAGTGCA141010AACTG CAGAACCAAT G CATTG G229090MAACTG 匚 AGCCAATGCATTGGAA2斗9090CTGCAGAACCAATGCATTGGATGCAT260PvulCCGATCGG8ATCGATCGAT101025TCGCGATCGCGA12010SacICGAG匚TC書81010SacIIGCCG匚GGC80TCC匚匸GCG石百GA125090SallGTCGACGTCAAAAGGCCATAGCGGCCGC2S00GCGTCGACGTCTTGGCCATAGCGGCCGCGG2010E0ACGCGTCGACGTCGGCCATAGCGGCCGCGGZA321075SealGAGTA tZTC81025AAAAGTAC iTTT12/b/hSmalCCCGGG6010CCCCGGGG8010CCCCCGGGGG101050TCGGCCGGGGGA129090SpelGACTAGTC810GGA匚TKGT匚匚101090CGGA匚TZGT匚CG12QEdCTAGA 匚 TMTCTAG14050SphIGG匚ATGCC8
