CLONING AND EXPRESSION OF SPODOPTERA LITURA UBIQUITIN GENE |
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Authors: | LI Zhao-fei GONG Ying-xue SHI Yong-xia PAN Li-jin PANG Yi |
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Institution: | State Key Laboratory-for Biocontrol, Zhongshan University, Guangzhou 510275, China |
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Abstract: | Abstract Ubiquitin (UBI) plays a very important role in regulated non-lysosomal ATP dependent protein degradation. In the present work, the coding sequence of Spodoptera litura UBI gene was isolated (GenBank Accession No. AF436066). The length of this ORF is 228bp, encoding a protein with Mr of 8.56 kD and isoelectric point of 6.56. Multiple sequence alignment indicated that S. litura UBI is very similar to the homologous proteins of other eukaryotic species and it has 84% identity with S. litura nucleopolyhedrovirus (SpltMNPV) UBI at amino acid level. RT-PCR analysis showed that S. litura UBI gene is ubiquitously expressed in larva tissues which are susceptible to SpltMNPV infection. By constructing E. coli expression vector, S. litura UBI was highly expressed and the recombinant protein was purified using Ni-NTA resin column, and currently further study on the function of S. litura UBI in SpltMNPV infection is underway. |
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Keywords: | Spodoptera litura ubiquitin gene cloning gene expression |
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