Purification, crystallization and preliminary X-ray crystallographic analysis of Pyrococcus furiosus DNA polymerase

被引：6

作者：

Goldman, S ^{[1
]}

Kim, R

Hung, LW

Jancarik, J

Kim, SH

机构：

[1] Univ Calif Berkeley, Lawrence Berkeley Lab, Phys Biosci Div, Berkeley, CA 94720 USA

[2] Univ Calif Berkeley, Dept Chem, Berkeley, CA 94720 USA

来源：

ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY | 1998年 / 54卷

关键词：

D O I：

10.1107/S0907444998000353

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

DNA polymerase gene from the hyperthermophilic Archaeon Pyrococcus furiosus has been cloned and the protein overexpressed in Escherichia coli to produce an active enzyme. The purified protein was crystallized from 0.08 M ammonium sulfate, 0.05 M Na-cacodylate, pH 6.5, 0.15%(v/v) NP40, 0.05%(v/v) Tween 20 and 4.5% (w/v) polyethylene glycol 6000 by the vapour-diffusion method. The orthorhombic crystals had unit-cell dimensions of a = 92.5, b = 125.4, c = 192.1 Angstrom: alpha = beta = gamma = 90 degrees. The crystals diffracted beyond 4 Angstrom on a 1.08 Angstrom synchrotron radiation source.

引用

页码：986 / 988

页数：3

共 16 条

[1]

BERGSEID M, 1992, STRATEGIES, V5, P50

[2]

Cline J., 1995, STRATEGIES MOL BIOL, V8, P24

[3]

CLINE JM, 1992, STRATEGIES, V4, P34

[4] AN ATTEMPT TO UNIFY THE STRUCTURE OF POLYMERASES [J].

DELARUE, M ;

POCH, O ;

TORDO, N ;

MORAS, D ;

ARGOS, P .

PROTEIN ENGINEERING, 1990, 3 (06) :461-467

[5] SPARSE-MATRIX SAMPLING - A SCREENING METHOD FOR CRYSTALLIZATION OF PROTEINS [J].

JANCARIK, J ;

KIM, SH .

JOURNAL OF APPLIED CRYSTALLOGRAPHY, 1991, 24 :409-411

[6] Crystal structure of a thermostable Bacillus DNA polymerase I large fragment at 2.1 angstrom resolution [J].

Kiefer, JR ;

Mao, C ;

Hansen, CJ ;

Basehore, SL ;

Hogrefe, HH ;

Braman, JC ;

Beese, LS .

STRUCTURE, 1997, 5 (01) :95-108

[7] CRYSTAL-STRUCTURE OF THERMUS-AQUATICUS DNA-POLYMERASE [J].

KIM, Y ;

EOM, SH ;

WANG, JM ;

LEE, DS ;

SUH, SW ;

STEITZ, TA .

NATURE, 1995, 376 (6541) :612-616

[8]

KONOLEV S, 1995, P NATL ACAD SCI USA, V92, P9264

[9] CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].

LAEMMLI, UK .

NATURE, 1970, 227 (5259) :680-+

[10] HIGH-FIDELITY AMPLIFICATION USING A THERMOSTABLE DNA-POLYMERASE ISOLATED FROM PYROCOCCUS-FURIOSUS [J].

LUNDBERG, KS ;

SHOEMAKER, DD ;

ADAMS, MWW ;

SHORT, JM ;

SORGE, JA ;

MATHUR, EJ .

GENE, 1991, 108 (01) :1-6

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