Cloning and Expression in Saccharomyces cerevisiae of a Synthetic Gene for the Type-I Trophoblast Interferon Ovine Trophoblast Protein-1: Purification and Antiviral Activity

Troy Ott, Gino Van Heeke, Howard M. Johnson, Fuller W. Bazer

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

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 × 108 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.

Original languageEnglish (US)
Pages (from-to)357-364
Number of pages8
JournalJournal of Interferon Research
Volume11
Issue number6
DOIs
StatePublished - Jan 1 1991

Fingerprint

Synthetic Genes
Interferon Type I
Trophoblasts
Antiviral Agents
Saccharomyces cerevisiae
Organism Cloning
Sheep
Yeasts
trophoblastin
Ion Exchange
Gel Chromatography
Plasmids
Mothers
Escherichia coli
Pregnancy
Mutation

All Science Journal Classification (ASJC) codes

  • Immunology
  • Virology

Cite this

@article{c431f3d1b3de4b8daf1b21109808eadd,
title = "Cloning and Expression in Saccharomyces cerevisiae of a Synthetic Gene for the Type-I Trophoblast Interferon Ovine Trophoblast Protein-1: Purification and Antiviral Activity",
abstract = "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 × 108 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.",
author = "Troy Ott and {Van Heeke}, Gino and Johnson, {Howard M.} and Bazer, {Fuller W.}",
year = "1991",
month = "1",
day = "1",
doi = "10.1089/jir.1991.11.357",
language = "English (US)",
volume = "11",
pages = "357--364",
journal = "Journal of Interferon Research",
issn = "0197-8357",
publisher = "Mary Ann Liebert Inc.",
number = "6",

}

Cloning and Expression in Saccharomyces cerevisiae of a Synthetic Gene for the Type-I Trophoblast Interferon Ovine Trophoblast Protein-1 : Purification and Antiviral Activity. / Ott, Troy; Van Heeke, Gino; Johnson, Howard M.; Bazer, Fuller W.

In: Journal of Interferon Research, Vol. 11, No. 6, 01.01.1991, p. 357-364.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cloning and Expression in Saccharomyces cerevisiae of a Synthetic Gene for the Type-I Trophoblast Interferon Ovine Trophoblast Protein-1

T2 - Purification and Antiviral Activity

AU - Ott, Troy

AU - Van Heeke, Gino

AU - Johnson, Howard M.

AU - Bazer, Fuller W.

PY - 1991/1/1

Y1 - 1991/1/1

N2 - 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 × 108 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.

AB - 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 × 108 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.

UR - http://www.scopus.com/inward/record.url?scp=0026365942&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026365942&partnerID=8YFLogxK

U2 - 10.1089/jir.1991.11.357

DO - 10.1089/jir.1991.11.357

M3 - Article

C2 - 1800584

AN - SCOPUS:0026365942

VL - 11

SP - 357

EP - 364

JO - Journal of Interferon Research

JF - Journal of Interferon Research

SN - 0197-8357

IS - 6

ER -