RT-PCR was carried out using cDNA prepared from antennae of day 0 females and degenerate primers designed from conserved sequences of Apis mellifera odorant receptor 2 (Or2), transcript variant X1, mRNA (gi 571501583) and Microplitis demolitor odorant receptor coreceptor (LOC103571567), mRNA (gi 665785588). Amplified fragments of A. japonica Orco gene were sequenced by using BigDye Terminator v3.1 (Applied Biosystems, USA) with 3130 Genetic Analyzer (Applied Biosystems, USA) [33 (link)]. Alignment of sequences was carried out on BioEdit Sequence Alignment Editor 7. 0. 9. 0. (Ibis Biosciences, USA).
To obtain full-length cDNA, 5’ rapid amplification of cDNA ends (5’RACE) was performed using 5’RACE kit (Invitrogen, Carlsbad, CA) as described previously [30 (link)]. All identified cDNA sequences were subjected to computer-assisted sequence analysis using GENETYX-MAC Ver. 13.1.7 (Software Development Co., Tokyo, Japan).
To obtain full-length cDNA, 5’ rapid amplification of cDNA ends (5’RACE) was performed using 5’RACE kit (Invitrogen, Carlsbad, CA) as described previously [30 (link)]. All identified cDNA sequences were subjected to computer-assisted sequence analysis using GENETYX-MAC Ver. 13.1.7 (Software Development Co., Tokyo, Japan).
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