EXPRESSION, PURIFICATION, AND FUNCTIONAL-CHARACTERIZATION OF ADENOVIRUS-5 AND ADENOVIRUS-12 E1A PROTEINS PRODUCED IN INSECT CELLS

被引：15

作者：

PEEPER, DS ^{[1
]}

ZANTEMA, A ^{[1
]}

DOWDY, SF ^{[1
]}

VANDEREB, AJ ^{[1
]}

机构：

[1] WHITEHEAD INST BIOMED RES,CAMBRIDGE,MA 02142

来源：

VIROLOGY | 1992年 / 190卷 / 02期

关键词：

D O I：

10.1016/0042-6822(92)90911-8

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

The 12 S and 13 S E1 A cDNAs from both the Adenovinus (Ad) nononcogenic type 5 and the oncogenic type 12 were overexpressed in an insect cell/baculovirus system. Upon infection of Spodoptera frugiperda cells, the production of E1A proteins reached a level of about 15 μg/106 Cells. The E1A proteins are highly soluble and apparently are processed authentically. They are readily recognized by various antibodies and display phosphorylation patterns similar to those of E1 A proteins synthesized in mammalian cells. Single-step immunoaffinity chromatography was used to purify the Ad5 E1A proteins to near homogeneity under nondenaturing conditions. The Ad5 and Ad12 E1 A proteins are able to form complexes with the retinoblastoma susceptibility gene product (Rb) and other cellular proteins. Interestingly, the presence of a cellular extract seems to be a prerequisite for association between highly purified E1A and Rb polypeptides. © 1992.

引用

页码：733 / 745

页数：13

共 69 条

[1] STRUCTURE OF ADENOVIRUS 2 EARLY MESSENGER-RNAS [J].