CHARACTERIZATION OF HUMAN DNA-LIGASE-I EXPRESSED IN A BACULOVIRUS-INSECT CELL SYSTEM

被引：4

作者：

GALLINA, A ^{[1
]}

ROSSI, F ^{[1
]}

MILANESI, G ^{[1
]}

ROSSI, R ^{[1
]}

MONTECUCCO, A ^{[1
]}

CIARROCCHI, G ^{[1
]}

机构：

[1] UNIV PAVIA,IST GENET BIOCHIM & EVOLUZIONIST,I-27100 PAVIA,ITALY

来源：

BIOCHEMICAL JOURNAL | 1995年 / 312卷

关键词：

D O I：

10.1042/bj3120593

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：

The baculovirus expression system used to overexpress human DNA ligase I (hLig I). Approx. 2 mg of recombinant hLig I was produced per 10(8) Spodoptera frugiperda Sf9 insect cells infected with the recombinant baculovirus. The optimum time point for the production of biologically active recombinant hLig I was 48 h post-infection. Lig I activity was demonstrated by auto-adenylating, polynucleotide joining and DNA relaxation assays. The baculovirus system has the advantage over previously described methods for producing hLig I of generating large amounts of a full-length active protein.

引用

页码：593 / 597

页数：5

共 20 条

[1] HUMAN DNA LIGASE-I CDNA - CLONING AND FUNCTIONAL EXPRESSION IN SACCHAROMYCES-CEREVISIAE [J].

BARNES, DE ;

JOHNSTON, LH ;

KODAMA, K ;

TOMKINSON, AE ;

LASKO, DD ;

LINDAHL, T .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (17) :6679-6683

[2]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[3] BACTERIOPHAGE-T4 AND HUMAN TYPE-I DNA LIGASES RELAX DNA UNDER JOINING CONDITIONS [J].

CIARROCCHI, G ;

LESTINGI, M ;

WRIGHT, G ;

MONTECUCCO, A .

NUCLEIC ACIDS RESEARCH, 1993, 21 (25) :5934-5939

[4] RAT-LIVER DNA LIGASES - CATALYTIC PROPERTIES OF A NOVEL FORM OF DNA-LIGASE [J].

ELDER, RH ;

MONTECUCCO, A ;

CIARROCCHI, G ;

ROSSIGNOL, JM .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1992, 203 (1-2) :53-58

[5]

ELDER RH, 1990, BIOCHEMISTRY-US, V29, P6010

[6]

FRASER MJ, 1992, CURR TOP MICROBIOL I, V158, P1

[7] INVITRO MUTAGENESIS AND FUNCTIONAL EXPRESSION IN ESCHERICHIA-COLI OF A CDNA-ENCODING THE CATALYTIC DOMAIN OF HUMAN DNA LIGASE-I [J].

KODAMA, K ;

BARNES, DE ;

LINDAHL, T .

NUCLEIC ACIDS RESEARCH, 1991, 19 (22) :6093-6099

[8]

LASKO DD, 1990, J BIOL CHEM, V265, P12618

[9] MAMMALIAN DNA LIGASES [J].

LINDAHL, T ;

BARNES, DE .

ANNUAL REVIEW OF BIOCHEMISTRY, 1992, 61 :251-281

[10]

MODRICH P, 1970, J BIOL CHEM, V245, P3626

← 1 2 →