Expression of two S-ribonucleases of Petunia inflata using baculovirus expression system

Biochem Biophys Res Commun. 1992 Aug 31;187(1):299-304. doi: 10.1016/s0006-291x(05)81492-5.

Abstract

We have previously shown that three Petunia inflata S-proteins, products of the multiallelic S-gene of the self-incompatibility system, are ribonucleases. Here we report the expression of cDNAs for two of these S-proteins using the baculovirus expression system. S2- and S3-proteins were found in both supernatants and lysates of Spodoptera frugiperda cells infected with recombinant baculoviruses. Both recombinant S-proteins contained glycosylated (25 kD) and nonglycosylated (23 kD) forms. Recombinant S2- and S3-proteins were purified from insect cell cultures, and the amino-terminal sequences determined from glycosylated S2- and S3-proteins indicated that the leader peptide encoded by each cDNA was correctly removed. Both glycosylated and nonglycosylated forms of S2- and S3-proteins exhibited ribonuclease activity.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Blotting, Western
  • Cell Line
  • DNA / genetics
  • Gene Expression*
  • Genetic Vectors
  • Glycosylation
  • Moths
  • Peptide Fragments / genetics*
  • Peptide Fragments / metabolism
  • Plants / enzymology*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Ribonucleases / genetics*
  • Ribonucleases / metabolism
  • Transfection

Substances

  • Peptide Fragments
  • Recombinant Proteins
  • ribonuclease S-peptide
  • DNA
  • Ribonucleases