期刊
INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
卷 31, 期 10, 页码 949-964出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/S0965-1748(01)00043-1
关键词
acyl-CoA desaturase; sex pheromone biosynthesis; lepidoptera; noctuidae; Helicoverpa zea; Trichoplusia ni; RT-PCR; cDNA; intron conservation; functional expression; Saccharomyces cerevisiae
In this report, we describe the structural and functional analyses of four acyl-CoA desaturase-encoding cDNAs that we isolated from RNA expressed in the pheromone gland of the com earworm, Helicoverpa zea. We deduced the homology relationships of the encoded proteins, designated HzPGDs1, HzPGDs2, HzPGDs3 and HzFBDs, to each other and to previously described desaturases of the cabbage looper moth, Trichoplusia ni, the fly, Drosophila melanogaster, and other more distantly related organisms. We also isolated genomic DNA fragments of the four H. zea desaturase-encoding genes, determined the locations of introns present in them, and compared them to conserved intron positions in reported desaturase genes of other species. We measured the levels of the four desaturase mRNAs in H. zea pheromone glands and larval fat bodies by RT-PCR. We established the functional identities of the deduced proteins HzPGDs1 and HzPGDs2, encoded by the two desaturase mRNAs that are differentially and abundantly expressed in pheromone glands of sexually mature adult H. Zea females, by functional expression of their encoding cDNAs in a desaturase-deficient mutant, olel, of the yeast Saccharomyces cerevisiae. We compared the unique unsaturated fatty acid profiles of HzPGDs1- and HzPGDs2-expressing transformants to those of strains expressing previously described Delta 11 and Delta9 desaturases of T ni. (C) 2001 Elsevier Science Ltd. All rights reserved.
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