Cloning and molecular characterization of a novel lectin gene from Pinellia ternata
| Autor: | Zhihua Liao, Xiu Yun Zhao, Juan Lin, Jun Song, Kexuan Tang, Fei Xavier Chen, Zhong Hai Chen, Jian Hong Yao, Xiao Fen Sun |
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| Rok vydání: | 2003 |
| Předmět: |
Models
Molecular Pinellia ternata Pinellia Recombinant Fusion Proteins Molecular Sequence Data Biology Protein Structure Secondary Plasmid Complementary DNA otorhinolaryngologic diseases Animals Gene family Amino Acid Sequence Northern blot Cloning Molecular Molecular Biology Gene Peptide sequence Southern blot Base Sequence Cell Biology biology.organism_classification Molecular biology Protein Structure Tertiary Aphids Plant Lectins Sequence Alignment |
| Zdroj: | Cell Research. 13:301-308 |
| ISSN: | 1748-7838 1001-0602 |
| DOI: | 10.1038/sj.cr.7290175 |
| Popis: | The full-length cDNA of Pinellia ternata agglutinin (PTA) was cloned from inflorescences using RACE-PCR. Through comparative analysis of PTA gene (pta) and its deduced amino acid sequence with those of other Araceae species, pta was found to encode a precursor lectin with signal peptide and to have extensive homology with those of other Araceae species. PTA was a heterotetrameric mannose-binding lectin with three mannose-binding boxes like lectins from other Araceae and Amaryllidaceae species. Southern blot analysis of the genomic DNA revealed that pta belonged to a low-copy gene family. Northern blot analysis demonstrated that pta constitutively expressed in various plant tissues including root, leaf, stem and inflorescence. The pta cDNA sequence encoding for mature PTA protein was cloned into pET-32a plasmid and the resulting plasmid, pET-32a-PTA containing Trx-PTA fusion protein, was investigated for the expression in E. coli BL21. SDS-PAGE gel analysis showed that the Trx-PTA fusion protein was successfully expressed in E. coli BL21 when induced by IPTG. Artificial diet assay revealed that PTA fusion protein had significant levels of resistance against peach potato aphids when incorporated into artificial diet at 0.1% (w/v). The cloning of the pta gene will enable us to further test its effect in depth on aphids by transferring the gene into crop plants. |
| Databáze: | OpenAIRE |
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