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FokI限制性内切酶的过量产生与结晶

Overproduction and crystallization of FokI restriction endonuclease.

作者信息

Kita K, Kotani H, Hiraoka N, Nakamura T, Yonaha K

机构信息

Central Research Laboratories, Takara Shuzo Co., Ltd., Shiga, Japan.

出版信息

Nucleic Acids Res. 1989 Nov 11;17(21):8741-53. doi: 10.1093/nar/17.21.8741.

DOI:10.1093/nar/17.21.8741
PMID:2685747
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC335040/
Abstract

To overproduce FokI endonuclease (R.FokI) in an Escherichia coli system, the coding region of R.FokI predicted from the nucleotide sequence was generated from the FokI operon and joined to the tac promoter of an expression vector, pKK223-3. By introduction of the plasmid into E. coli UT481 cells expressing the FokI methylase gene, the R.FokI activity was overproduced about 30-fold, from which R.FokI was purified in amounts sufficient for crystallization. The removal of a stem-loop structure immediately upstream of the R.FokI coding region was essential for overproduction.

摘要

为了在大肠杆菌系统中过量表达FokI核酸内切酶（R.FokI），根据核苷酸序列预测的R.FokI编码区从FokI操纵子中获得，并连接到表达载体pKK223-3的tac启动子上。通过将该质粒导入表达FokI甲基化酶基因的大肠杆菌UT481细胞中，R.FokI活性过量表达了约30倍，从中纯化出了足以用于结晶的R.FokI。去除R.FokI编码区上游紧邻的茎环结构对于过量表达至关重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a93d/335040/7eb1210a5ce6/nar00138-0344-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a93d/335040/e83ddcccbc03/nar00138-0339-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a93d/335040/779226edb534/nar00138-0342-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a93d/335040/669f4e4073c8/nar00138-0343-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a93d/335040/7eb1210a5ce6/nar00138-0344-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a93d/335040/e83ddcccbc03/nar00138-0339-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a93d/335040/779226edb534/nar00138-0342-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a93d/335040/669f4e4073c8/nar00138-0343-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a93d/335040/7eb1210a5ce6/nar00138-0344-a.jpg

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本文引用的文献

1
Characterization of the genes coding for the Eco RV restriction and modification system of Escherichia coli.大肠杆菌Eco RV限制与修饰系统编码基因的特性分析。
Nucleic Acids Res. 1984 Apr 25;12(8):3659-76. doi: 10.1093/nar/12.8.3659.
2
New restriction endonucleases from Flavobacterium okeanokoites (FokI) and Micrococcus luteus (MluI).来自海洋黄杆菌（FokI）和藤黄微球菌（MluI）的新型限制性内切酶。
Gene. 1981 Dec;16(1-3):73-8. doi: 10.1016/0378-1119(81)90062-7.
3
Isolation of gram quantities of EcoRI restriction and modification enzymes from an overproducing strain.
乳球菌质粒编码的II型限制/修饰系统LlaDII的克隆与特性分析
Appl Environ Microbiol. 1998 Jul;64(7):2424-31. doi: 10.1128/AEM.64.7.2424-2431.1998.
4
Alteration of the cleavage distance of Fok I restriction endonuclease by insertion mutagenesis.通过插入诱变改变福克I型限制性内切酶的切割距离。
Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2764-8. doi: 10.1073/pnas.90.7.2764.
5
Purification and properties of the MboII, a class-IIS restriction endonuclease.IIS类限制性内切酶MboII的纯化及特性
Nucleic Acids Res. 1992 Feb 11;20(3):433-8. doi: 10.1093/nar/20.3.433.
6
Functional domains in Fok I restriction endonuclease.福克I型限制性内切核酸酶中的功能结构域。
Proc Natl Acad Sci U S A. 1992 May 15;89(10):4275-9. doi: 10.1073/pnas.89.10.4275.
7
Cloning and sequence analysis of the StsI restriction-modification gene: presence of homology to FokI restriction-modification enzymes.StsI 限制修饰基因的克隆与序列分析：与 FokI 限制修饰酶存在同源性
Nucleic Acids Res. 1992 Aug 25;20(16):4167-72. doi: 10.1093/nar/20.16.4167.
从高产菌株中分离克级量的EcoRI限制酶和甲基化酶。
J Biol Chem. 1984 Sep 25;259(18):11571-5.
4
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.在噬菌体T4头部组装过程中结构蛋白的切割
Nature. 1970 Aug 15;227(5259):680-5. doi: 10.1038/227680a0.
5
Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes.从电转印到聚偏二氟乙烯膜上的皮摩尔量蛋白质中获得的序列。
J Biol Chem. 1987 Jul 25;262(21):10035-8.
6
The nucleotide sequence of pACYC184.pACYC184的核苷酸序列。
Nucleic Acids Res. 1988 Jan 11;16(1):355. doi: 10.1093/nar/16.1.355.
7
Nucleotide sequence of the lig gene and primary structure of DNA ligase of Escherichia coli.大肠杆菌连接基因的核苷酸序列及DNA连接酶的一级结构
Mol Gen Genet. 1986 Jul;204(1):1-7. doi: 10.1007/BF00330179.
8
Universal restriction endonucleases: designing novel cleavage specificities by combining adapter oligodeoxynucleotide and enzyme moieties.通用限制性核酸内切酶：通过组合衔接子寡脱氧核苷酸和酶部分来设计新型切割特异性
Gene. 1985;40(2-3):169-73. doi: 10.1016/0378-1119(85)90039-3.
9
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J Biol Chem. 1985 Dec 5;260(28):15339-44.
10
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Gene. 1985;37(1-3):229-39. doi: 10.1016/0378-1119(85)90277-x.