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| Molecular Cloning of Ubiquitin Extension Protein Gene of Helicoverpa assulta and Its Expression in E. coli |
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Abstract The cDNA encoding the ubiquitin-53aa extension protein(ubi-53,UBE) was isolated from the fat of Helicoverpa assulta larvae by reverse transcription polymerase chain reaction (RT-PCR). The UBE in H. assulta was 390bp in length, encoded a peptide of 129 amino acid residues, in which there was an ubiquitin fused with a ribosomal L40 protein, the predicted MW was 14.8 kDa. The deduced amino acid sequence has a high identity (90%-98%) with the reported sequence of UBE from other eukaryotic species and 69% with H. armigera single nucleopolyhedrovirus (HaSNPV) ubiquitin. The fragment containing UBE gene was inserted into pGEX-4T-2 expressive vector, and the expression was induced by IPTG in E. coli BL21(DE3). It’s molecular weight was about 40 kDa, by checking with SDS polyacrylamide gel electrophoresis and Western blotting using a mouse monoclonal antibody against GST.
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Received: 21 March 2006
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