Genomic cloning, bioinformatics analysis, and subcellular localization of an agglutinin gene from Pinellia ternate / 中草药

ABSTRACT

Objective: To clone an agglutinin gene from Pinellia ternata and to analyze its bioinformatics and subcellular location. Methods: Based on the published sequence GU593718.1 from Genbank, P. ternata agglutinin (PTA) was amplified and cloned from genomic DNA of the fresh leaves of P. ternata. The cloned PTA gene was further fused to the plant expression vector pI1300-CaMV35S-GFP to construct pI1300-CaMV35S-PTA-GFP, then transfered into cells of Agrobacterium tumefaciens GV3101. Its transient expression was observed in Nicotiana tabacum. Results: The full length of PTA contained 810 bp with the deduced 269 amino acid residues; It contained one signal peptide, two conversation B-lectin domains and three mannose binding sites; PTA shared 97%, 85%, and 83% identity with the amino acid sequence from PTA, and Pinellia pedatisecta agglutinin (PPA), Pinellia cordata agglutinin (PCA), respectively; The PTA was localized to the plasma membrane; Its registration number is KF154979 in NCBI. Conclusion: It would provide a stable foundation for the study on its effect against fungi, insects, and bacterium.