Expression of two S-ribonucleases of Petunia inflata using baculovirus expression system.
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.
Mu
J
J
Department of Molecular and Cell Biology, Pennsylvania State University, University Park 16802.
Kao
T H
TH
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D002460:Cell Line; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006031:Glycosylation; D009036:Moths; D010446:Peptide Fragments / Q000235:genetics* / Q000378:metabolism; D010944:Plants / Q000201:enzymology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012260:Ribonucleases / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
D010446:Peptide Fragments; D011994:Recombinant Proteins; C023560:ribonuclease S-peptide; D004247:DNA; D012260:Ribonucleases