Construction of fulllength cDNA library of winterdiapause pupae of the onion maggot, Delia antique.
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Key Words：Delia antiqua, winterdiapause pupae, fulllength cDNA library, SMART technique
Abstract： The onion maggot, Delia antiqua（Meigen）, is characterized by winter-diapause and is close to Drosophila melanogaster in phelogenetics. This study aims to construct full-length cDNA library of winterdiapause pupae of D. antique, in order to provide a base for further screening, cloning and expression analysis of winter-diapausespecific genes. In this study, total RNA was extracted from winterdiapause pupae of D. antiqua using RNAiso kit. Double-stranded cDNAs were synthesized with SMART technique and digested by the enzyme SfiI, and then the cDNAs were ligated into the vector pDNR-LIB. The ligation mixture was transformed into E. coli DH10B by eletroporation. According to the evaluation on quality, the titer of primary library was 2.4×107 cfu/mL. The results from random sampling of 15 clones showed that the inserted fragments ranged from 0.5 kb to 3 kb by PCR amplification, with an average size of 1 kb, and the recombination rate was 100 %. These results showed that a full-length cDNA library with high quality on D. antiqua winter-diapause pupae was well constructed.