PRODUCTION OF ACTIVE-MOUSE DNA-POLYMERASE-DELTA IN BACTERIA

被引：22

作者：

HINDGES, R ^{[1
]}

HUBSCHER, U ^{[1
]}

机构：

[1] UNIV ZURICH,INST VET BIOCHEM,CH-8057 ZURICH,SWITZERLAND

来源：

GENE | 1995年 / 158卷 / 02期

关键词：

DNA REPLICATION; RECOMBINANT DNA; T7-MEDIATED OVEREXPRESSION; ANTIBODY; PCR;

D O I：

10.1016/0378-1119(95)00065-E

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

The entire cDNA encoding the large subunit of mouse DNA polymerase delta (mPol delta; EC 2.7.7.7) has been cloned and expressed in various bacterial expression systems. A soluble protein could only be obtained when mPol delta was produced as a glutathione S-transferase (GST) fusion protein and the incubation temperature of the expression strain was reduced to 30 degrees C. After purification over a glutathione-Sepharose column, the fractions containing the recombinant (re-) fusion protein showed both DNA Pol and 3'-->5' fro activities. In situ activity gel analysis indicated that the Pol activity resides in the re-protein. This activity, however, was not stimulated by proliferating cell nuclear antigen (PCNA). Our data are discussed in the view of the findings of Goulian et al. [J. Biol. Chem., 265 (1990) 16402-16411] that the second mPol delta subunit, the 48-kDa protein, might play an important role in DNA Pol delta-PCNA interaction.

引用

页码：241 / 246

页数：6

共 21 条

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