DNA SPECIFICITY OF THE CRE RECOMBINASE RESIDES IN THE 25 KDA CARBOXYL DOMAIN OF THE PROTEIN
DNA SPECIFICITY OF THE CRE RECOMBINASE RESIDES IN THE 25 KDA CARBOXYL DOMAIN OF THE PROTEIN
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DOI：
10.1016/s0022-2836(99)80007-2
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发表时间：
1990-12-20
影响因子：
5.6
通讯作者：
MACK, A
中科院分区：
文献类型：
--
作者：
HOESS, R;ABREMSKI, K;MACK, A
噬菌体P1的Cre蛋白是一种38.5 kDa的位点特异性重组酶，属于Int重组蛋白家族。Cre通过与34个碱基对的序列lox特异性结合来发挥作用，在那里它进行重组。有限的Cre胰凝乳酶消化产生了两个大小分别为25 kDa和13.5 kDa的片段。PURIFID 25 kDa多肽的氨基末端序列表明，该多肽代表Cre蛋白的羧基末端部分。构建了cre基因的截短版本，它只产生25 kDa的多肽。25 kDa的多肽能够与lox位点特异结合，但结合亲和力低于野生型Cre。Fe-EDTA的足迹显示，25 kDa的多肽保护lox位点的反向重复序列，但仅对间隔区有部分保护。这与野生型Cre获得的足迹形成对比，野生型Cre保护整个间隔区。
The Cre protein of bacteriophage P1 is a 38.5 kDa site-specific recombinase that belongs to the Int family of recombination proteins. Cre acts by binding specifically to a 34 base-pair sequence, lox, where it carries out recombination. A limited chymotryptic digest of Cre resulted in two fragments of sizes 25 and 13.5 kDa, respectively. The sequence of the amino terminus of the purifid 25 kDa peptide demonstrated that this peptide represents the carboxyl-terminal portion of the Cre protein. A truncated version of the cre gene was constructed which produces only the 25 kDa peptide. The 25 kDa peptide is capable of specific binding to the lox site, but binds at lower affinity than does wild-type Cre. Footprinting with Fe-EDTA indicates that the 25 kDa peptide protects the inverted repeats of the lox site but shows only partial protection of the spacer region. This is in contrast to the footprint obtained with wild-type Cre which protects the entire spacer region.