Isolation of white gene orthologue of the sawfly, Athalia rosae (Hymenoptera) and its functional analysis using RNA interference.
We isolated and characterized the white gene orthologue of the sawfly, Athalia rosae (Hymenoptera). The A. rosae white (Ar white) cDNA cloned was 2058-bp long encoding 685 amino acids in a single open reading frame (ORF). Comparison of the cDNA sequence with the genomic DNA sequence revealed that the ORF was derived from 11 exons. Ar white was a single copy gene as evidenced by genomic Southern blotting and its cytological localization on the metaphase chromosomes. The deduced amino acid sequence aligned well with known insect white orthologous gene products sharing conserved regions such as the ATP-binding motif and the six transmembrane-spanning segments. Expression of Ar white was detected at embryonic and pupal stages by Northern blotting. In situ hybridization detected the embryonic expression in a pair of the lateral tips of protocephalic placodes from where optic organs are formed. Ar white function was examined using double-stranded RNA (dsRNA)-mediated interference. The synthesized dsRNA targeting Ar white transcripts caused a decrease in the level of the original mRNAs, and resulted in the white phenocopy in the embryonic eye pigmentation when microinjected into eggs from wild-type females. The effects occurred in a dose-dependent manner.
Sumitani
Megumi
M
Division of Bioscience, Graduate School of Science and Technology, Kobe University, Nada, Kobe 657-8501, Japan.
Yamamoto
Daisuke S
DS
Lee
Jae Min
JM
Hatakeyama
Masatsugu
M
D018528:ATP-Binding Cassette Transporters / Q000235:genetics* / Q000378:metabolism; D020816:Amino Acid Motifs / Q000235:genetics; D000818:Animals; D003001:Cloning, Molecular; D018507:Gene Expression Regulation, Developmental / Q000235:genetics* / Q000502:physiology; D006927:Hymenoptera / Q000196:embryology / Q000235:genetics*; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*
D018528:ATP-Binding Cassette Transporters; D019476:Insect Proteins; D034741:RNA, Small Interfering