| Products/Services Used | Details | Operation |
|---|---|---|
| Peptide Synthesis | A construction coupling the P. vulgaris polygalacturonase (PGIP) signal peptide at the 5´-end, and 6xHis-tail at the 3´-end of the Tepary bean lectin gene was synthesized by GenScript USA Inc. Once the construction was obtained, it was released from the commercial vector pUC57 by digestion carried out with the BamHI restriction enzyme (Fermentas, Ontario, Canada), and incubated at 37 °C from 3 to 5 h. The sequence was introduced in a previously digested vector pBI121, by a ligation carried out with the T4 ligase (Fermentas, Ontario, Canada), incubating at 16 °C for 16 h. Subsequently, the result of the previous ligation was cloned into chemocompetent cells of E. coli Top 10 by heat shock (Sambrook et al., 1989). The transformed colonies were recovered for growth on plates of LB medium with 100 μg/mL of ampicillin as selection agent, incubated at 37 °C for 24 h, and finally the plasmids were extracted by the miniprep method (Birnboim and Doly, 1979). | Get A Quote |
Tepary bean (Phaseolus acutifolius) lectin fraction (TBLF) has been shown to specifically bind and induce cell death of different types of cancer cells and also has exhibited an effect on early colon tumorigenesis. However, the development of a pharmaceutical formula is not possible yet because the production process is expensive and slow and provides low yields. Therefore, the purpose of the present work was to develop a strategy to produce one bioactive lectin by rhizosecretion through root exudates on genetically modified plants. Amplification of Tepary bean transcripts was performed using degenerate primers, and the products obtained were sequenced. Multiple alignments of sequences led to elucidating one of... More
