Isolation, Target Site Prediction and Validation of miR393 Prediction Target Gene Ve in Solanum torvum
Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, 210014
Bioscience Methods, 2020, Vol. 11, No. 1 doi: 10.5376/bm.2020.11.0001
Received: 01 Dec., 2020 Accepted: 04 Dec., 2020 Published: 11 Dec., 2020
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Preferred citation for this article:
Qin Y.L., Wang Z., Shu X.C., Zhuang W.B., Zhang F.J., and Peng F., 2020, Isolation, target site prediction and validation of miR393 prediction target gene Ve in Solanum torvum, Bioscience Method, 11(1): 1-9 (doi: 10.5376/bm.2020.11.0001)
In this study, total RNA was extracted by modified Triziol method and the full-length sequence of Verticillium wilt resistance gene Ve was obtained by homologous cloning. The binding site of miR393 to Ve was predicted by rna hybrid and validated by RLM-5’-RACE. The results showed that Ve gene contains 3302 nucleotides, encodes 904 amino acids, and has 85.63% homology with tomato, potato, tobacco, etc. The predicted results of binding sites showed that miR393 and Ve were almost completely paired from site 263 in Solanum torvum. With the help of RLM-5’-RACE, we found that the miR393 and target gene Ve were cut between the 10th and 11th base sites of the complementary binding sequence. Through the verification of PCR and sequencing, Ve was finally confirmed as the target gene of miR393 in Solanum torvum. It provides theoretical basis for further research on the molecular mechanism of miR393 mediated Ve gene participating in Verticillium wilt resistance.
Solanum torvum Swartz; Target gene; miR393; Ve gene; Verticillium wilt