|Perera, O. - UNIVERSITY OF FLORIDA|
Submitted to: Archives of Insect Biochemistry and Physiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 12, 1999
Publication Date: N/A
Interpretive Summary: The expected loss of methyl bromide as a fumigant, as well as some other insecticides because of acquired resistance or environmental hazard will leave the stored product industry with fewer options for pest control. Scientists at the USDA, ARS, Center for Medical, Agricultural and Veterinary Entomology, in Gainesville, FL, are developing genetic and physiological based methods of control for integrated pest management programs. To find a method to produce sterile insects, the gene for a yolk protein (YP4) produced within the ovary was identified and isolated from the Indianmeal moth. The gene sequence is similar only to a gene for a related protein in the greater wax moth. DNA regulatory sequences for this gene will be used to develop methods to genetically sterilize the Indianmeal moth.
Technical Abstract: YP4, a subunit of the follicular epithelium yolk protein in the moth, Plodia interpunctella, is produced in the follicle cells during vitellogenesis and after secretion is taken up into the oocyte and stored in the yolk spheres for utilization during embryogenesis. In order to identify the cDNA clones for YP4, a degenerate PCR primer was designed to six amino acid residues identified in the NH2-terminal sequence of mature YP4. In conjunction with the T7 reverse PCR primer from the Lambda Zap II vector, a PCR product for the majority of the coding sequence was generated. The 5'region of the YP4 transcript was determined by 5'RACE based on nested primers designed from the YP4 cDNA sequence. The combined cDNA and 5'RACE sequencing showed the YP4 transcript to be 991 bp in length with a single open reading frame for a predicted polypeptide of 299 amino acids. Northern analysis showed a single YP4 transcript was present in ovarian RNA that was approximately 1 kb in length. The predicted amino acid sequence for YP4 from P. interpunctella was most closely related to the predicted YP4 protein from the moth, Galleria mellonella, and the spherulin 2a protein from the slime mold, Physarum polycephalum.