We report the molecular cloning and nucleotide sequence of the nuclear gene, POX18, encoding an oleate-inducible peroxisomal protein from the yeast Candida tropicalis. POX18 has a single open reading frame of 381 nucleotides (nt), which encodes a protein of 127 amino acids. The predicted Mr of this protein is 13 792. Codon usage in the expression of POX18 is non-random, and shows a pattern similar to that used for other peroxisomal genes from C. tropicalis and highly expressed genes from Saccharomyces cerevisiae. Northern analysis of total RNA from oleate-grown cells determined that POX18 mRNA is approximately 750 nt in length. The POX18 gene was expressed in vitro, which resulted in a single translation product that co-migrated in denaturing polyacrylamide gels with an abundant peroxisomal protein (apparent mass of 16 kDa) and was immunoprecipitated by an antiserum against peroxisomal protein.
Bibliographical noteFunding Information:
We gratefully acknowledge Mr. Gary Richo, Mr. David Allman and Mr. Bruce Fieldman for technical assistance, and Drs. Gary Otto and Anthony Po-powics for assistance with the computer analysis. This work was supported by National Science Foundation grant DCB83-09222 and National Institute of Health grant DK-39684.
- Recombinant DNA
- gene structure
- hydropathy plot
- in vitro expression