Acta Entomologica Sinica (2004) 47, 166-170

Guo-Dong Niu, Hai-Yuan Zhang and Zhong-Xin Zhang (2004)
Amplication with 3' RACE-PCR , cloning and prokaryotic expression of ubiquitin extension protein cDNA of Spodoptera exigua (Lepidoptera : Noctuidae)
Acta Entomologica Sinica 47 (2), 166-170
Abstract: To identify the nucleotide sequence of ubiquitin cDNA in Spodoptera exigua, a 513 by cDNA fragment encoding ubiquitin-53aa extension protein from S. exigua lipid was amplified with 3'RACE (rapid amplification of cDNA ends) PCR with primers designed on the N-terminal amino acid sequence of S. frugiperda ubiquitin extension gene, and the amplified fragment was cloned and sequenced. The ubiquitin-53aa extension gene (ubi-53) in S. exigua was 513 bp in length, including 123 bp of 3' untranslated region and 390 bp of encoding region. The encoding region encoded a peptide of 129 amino acid residues, in which there was an ubiquitin fused with a ribosomal L40 protein. A comparison of the deduced amino acid sequence with those of Bombyx morn, S. frugiperda, Drosophila melanogaster, Homo sapienes , SeNPV and BmNPV showed that the amino acid homology rates were 96.9%, 98.5% , 95.3%, 93.0%, 78.8% and 77.2% respectively. This suggests ube genes in eukaryotes may have a different evolution way from its host viruses. The fragment containing ubi-53 gene was inserted into pET-28a expressive vector, and the expression was induced by IPTG in E . coli BL21 ( DE3) . The fusion protein was identified by Western blot using a mouse monoclonal antibody against bovine ubiquitin.
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Research topic(s) for pests/diseases/weeds:
molecular biology - genes

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Spodoptera exigua