Cloning of EF-1a and AK Genes from Mythimna separata and Establishment of a Real Time RT-PCR Assay

doi:10.11924/j.issn.1000-6850.casb15120120

Abstract

Abstract: In order to analyze the expression patterns of EF- 1a and AK genes in Mythimna separata by real time RT-PCR, the partial cDNAs of the two genes were cloned using the designed primers, and the sequence homology was analyzed. Then specific primers were designed and the real time RT- PCR method was established by analyzing melting curve and standard curve. Results showed the cloned cDNAs of EF-1a and AK genes were 622 bp and 1020 bp, and encoded peptides of 207 amino acids and 339 amino acids, respectively. The amino acid sequence of EF-1a shared homologies of above 95% with Bombyx mori, Biston betularia and Papilio xuthus, and the amino acid sequence of AK shared homologies of above 98% with Spodoptera frugiperda, Heliothis virescens, Helicoverpa armigera, Spodoptera litura and Spodoptera exigua. Analyses of melting curve and standard curve indicated that the designed primers of these two genes could be applied to real time RT-PCR experiments. This study could provide a foundation for future gene expression studies in M. separata.

CLC Number:

Q966

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