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Titolo:
CLONING AND SEQUENCE-ANALYSIS OF THE DNA LIGASE-ENCODING GENE OF RHODOTHERMUS-MARINUS, AND OVERPRODUCTION, PURIFICATION AND CHARACTERIZATION OF 2 THERMOPHILIC DNA LIGASES
Autore:
THORBJARNARDOTTIR SH; JONSSON ZO; ANDRESSON OS; KRISTJANSSON JK; EGGERTSSON G; PALSDOTTIR A;
Indirizzi:
UNIV ZURICH,INST VET BIOCHEM,WINTERTHURERSTR 190 CH-8057 ZURICH SWITZERLAND UNIV ICELAND,INST BIOL IS-108 REYKJAVIK ICELAND INST EXPTL PATHOL IS-128 REYKJAVIK ICELAND TECHNOL INST ICELAND IS-112 REYKJAVIK ICELAND
Titolo Testata:
Gene
fascicolo: 1, volume: 161, anno: 1995,
pagine: 1 - 6
SICI:
0378-1119(1995)161:1<1:CASOTD>2.0.ZU;2-N
Fonte:
ISI
Lingua:
ENG
Soggetto:
NUCLEOTIDE-SEQUENCE; MOLECULAR CHARACTERIZATION; ESCHERICHIA-COLI; EXPRESSION; PROTEIN;
Keywords:
NAD(+)-DEPENDENT DNA LIGASE; THERMOPHILIC BACTERIUM; COMPLEMENTATION; RECOMBINANT DNA; T7 OVEREXPRESSION; DNA REPLICATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
35
Recensione:
Indirizzi per estratti:
Citazione:
S.H. Thorbjarnardottir et al., "CLONING AND SEQUENCE-ANALYSIS OF THE DNA LIGASE-ENCODING GENE OF RHODOTHERMUS-MARINUS, AND OVERPRODUCTION, PURIFICATION AND CHARACTERIZATION OF 2 THERMOPHILIC DNA LIGASES", Gene, 161(1), 1995, pp. 1-6

Abstract

In this paper we describe the cloning and sequence analysis of a geneencoding DNA ligase (Lig; EC 6.5.1.2) from the thermophilic bacteriumRhodothermus marinus (Rm). We also describe the overexpression of theLig-encoding genes of Rm and the thermophile, Thermus scotoductus (Ts), in Escherichia coli, and the purification and characterization of the overproduced Lig. The Rm lig gene encodes a protein of 712 amino acids (aa) with a calculated molecular mass of 79 487 Da. Comparison with published sequences of bacterial Lig revealed significant homology between the NAD(+)-utilizing Lig, and alignment of their aa sequences revealed several blocks of conserved residues. Both of the purified Ligexhibit nick-closing activity over a wide range of temperatures. Under our assay conditions the Rm Lig was active at 5-75 degrees C with apparent optimal activity above 55 degrees C. The Ts enzyme showed activity at 15-75 degrees C with optimal activity above 65 degrees C. The half-life of the Lig at 91 degrees C was estimated to be 7 min for the Rm Lig and 26 min for the Ts Lig.

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Documento generato il 26/09/20 alle ore 01:08:04