An in vivo selection system for homing endonuclease activity -: art. no. e29

被引：23

作者：

Gruen, M ^{[1
]}

Chang, K ^{[1
]}

Serbanescu, I ^{[1
]}

Liu, DR ^{[1
]}

机构：

[1] Harvard Univ, Dept Chem & Biol Chem, Cambridge, MA 02138 USA

来源：

NUCLEIC ACIDS RESEARCH | 2002年 / 30卷 / 07期

基金：

美国国家科学基金会;

关键词：

D O I：

10.1093/nar/30.7.e29

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Homing endonucleases are enzymes that catalyze the highly sequence-specific cleavage of DNA. We have developed an in vivo selection in Escherichia coli that links cell survival with homing endonuclease-mediated DNA cleavage activity and sequence specificity. Using this selection, wild-type and mutant variants of three homing endonucleases were characterized without requiring protein purification and in vitro analysis. This selection system may facilitate the study of sequence-specific DNA cleaving enzymes, and selections based on this work may enable the evolution of homing endonucleases with novel activities or specificities.

引用

页数：6

共 28 条

[1] Active barnase variants with completely random hydrophobic cores [J].

Axe, DD ;

Foster, NW ;

Fersht, AR .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (11) :5590-5594

[2] Homing endonucleases: keeping the house in order [J].

Belfort, M ;

Roberts, RJ .

NUCLEIC ACIDS RESEARCH, 1997, 25 (17) :3379-3388

[3] Chemical probing shows that the intron-encoded endonuclease I-SceI distorts DNA through binding in monomeric form to its homing site [J].

Beylot, B ;

Spassky, A .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (27) :25243-25253

[4] Homing endonucleases: structural and functional insight into the catalysts of intron/intein mobility [J].

Chevalier, BS ;

Stoddard, BL .

NUCLEIC ACIDS RESEARCH, 2001, 29 (18) :3757-3774

[5] The monomeric homing endonuclease PI-SceI has two catalytic centres for cleavage of the two strands of its DNA substrate [J].

Christ, F ;

Schoettler, S ;

Wende, W ;

Steuer, S ;

Pingoud, A ;

Pingoud, V .

EMBO JOURNAL, 1999, 18 (24) :6908-6916

[6] RECOGNITION AND CLEAVAGE SITE OF THE INTRON-ENCODED OMEGA TRANSPOSASE [J].

COLLEAUX, L ;

DAURIOL, L ;

GALIBERT, F ;

DUJON, B .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (16) :6022-6026

[7] TIGHT REGULATION, MODULATION, AND HIGH-LEVEL EXPRESSION BY VECTORS CONTAINING THE ARABINOSE P-BAD PROMOTER [J].

GUZMAN, LM ;

BELIN, D ;

CARSON, MJ ;

BECKWITH, J .

JOURNAL OF BACTERIOLOGY, 1995, 177 (14) :4121-4130

[8] BARNASE AND BARSTAR - 2 SMALL PROTEINS TO FOLD AND FIT TOGETHER [J].

HARTLEY, RW .

TRENDS IN BIOCHEMICAL SCIENCES, 1989, 14 (11) :450-454

[9] BARNASE AND BARSTAR - EXPRESSION OF ITS CLONED INHIBITOR PERMITS EXPRESSION OF A CLONED RIBONUCLEASE [J].

HARTLEY, RW .

JOURNAL OF MOLECULAR BIOLOGY, 1988, 202 (04) :913-915

[10] Amino acid residues in both the protein splicing and endonuclease domains of the PI-SceI intein mediate DNA binding [J].

He, ZN ;

Crist, M ;

Yen, HC ;

Duan, XQ ;

Quiocho, FA ;

Gimble, FS .

JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (08) :4607-4615

← 1 2 3 →