2024年4月23日发(作者：win7系统关闭445端口)

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，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 produce 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 systems forming

either C5-methylcytosine, N4-methylcytosine or N6-methyladenine.

酶

ApaI

BamHI

BglII

类型

Type

enzyme

Type

enzyme

Type

enzyme

Type

enzyme

Type

enzyme

II restriction

II restriction

II restriction

II restriction

II restriction

识别序列

5'GGGCC^C 3'

5' G^GATCC 3'

5' A^GATCT 3'

EcoRI

HindIII

5' G^AATTC 3'

5' A^AGCTT 3'

KpnI

Type

enzyme

Type

enzyme

Type

enzyme

Type

enzyme

Type

enzyme

Type

enzyme

Type

enzyme

Type

enzyme

Type

enzyme

Type

enzyme

II restriction

5' GGTAC^C 3'

NcoI

II restriction

5' C^CATGG 3'

NdeI

II restriction

5' CA^TATG 3'

NheI

II restriction

5' G^CTAGC 3'

NotI

II restriction

5' GC^GGCCGC 3'

SacI

II restriction

5' GAGCT^C 3'

SalI

II restriction

5' G^TCGAC 3'

SphI

II restriction

5' GCATG^C 3'

XbaI

II restriction

5' T^CTAGA 3'

XhoI

II restriction

5' C^TCGAG 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 Premier5.0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：/rebase/）

当你设计好引物，添加上了内切酶识别序列，下一步或许是添加保护碱基了，可以参考：

NEB公司网站提供的关于设计PCR引物保护碱基参考表下载（也可见图片）