CLONING AND EXPRESSION IN SACCHAROMYCES-CEREVISIAE OF A SYNTHETIC GENE FOR THE TYPE-I TROPHOBLAST INTERFERON OVINE TROPHOBLAST PROTEIN-1 - PURIFICATION AND ANTIVIRAL ACTIVITY

被引:79
作者
OTT, TL
VANHEEKE, G
JOHNSON, HM
BAZER, FW
机构
[1] UNIV FLORIDA,DEPT ANIM SCI,BLDG 459,SHEALY DR,GAINESVILLE,FL 32611
[2] UNIV FLORIDA,INTERDISCIPLINARY CTR BIOTECHNOL RES,PROT EXPRESS LAB,GAINESVILLE,FL 32611
[3] UNIV FLORIDA,DEPT MICROBIOL & CELL SCI,GAINESVILLE,FL 32611
来源
JOURNAL OF INTERFERON RESEARCH | 1991年 / 11卷 / 06期
关键词
D O I
10.1089/jir.1991.11.357
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
Ovine trophoblast protein-1 (oTP-1) is a unique, Type I, trophoblast interferon (IFN) that possesses potent antiviral activity and is thought to be primarily responsible for maternal recognition of pregnancy in sheep. To provide sufficient amounts of protein for detailed studies, a synthetic gene for oTP-1 was designed and assembled in Escherichia coli, subcloned into a yeast expression plasmid, and used to overproduce recombinant oTP-1 in Saccharomyces cerevisiae. Recombinant oTP-1 was purified from soluble yeast extract using sequential ion-exchange and molecular sieve chromatography. Recombinant oTP-1 purified in this fashion exhibited potent antiviral activity (0.6 x 10(8) U/mg) similar to native oTP-1. This expression system will enable production of large quantities of soluble, biologically active, and correctly processed recombinant oTP-1. Furthermore, the synthetic gene construct facilitates introduction of mutations for ongoing structure/function studies of this unique, Type I, trophoblast IFN.
引用
收藏
页码:357 / 364
页数:8
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