Salt resistance gene StNHX1 of wild eggplant and herbicide resistance plant expression vector thereof
A plant expression vector, wild eggplant technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., to achieve the effect of improving salt tolerance and broad application prospects
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Embodiment 1
[0039] Example 1 Cloning of wild eggplant salt tolerance gene StNHX1
[0040] Wild eggplant 'Torvum Vigor' (Solanum torvum Swartz; Japanese rootstock variety, purchased from Japan Takii Seedling Co., Ltd.) was selected as the test material. When the seedlings grew to eight true leaves, 100 mmol L -1 After NaCl stress treatment for 3 days, young leaves were taken, and total RNA was extracted according to the instructions of the Total RNA Extraction Kit (TaKaRa), and 2 μg of total RNA was reverse-transcribed into cDNA (ReverTra Ace-a- TM cDNA synthesis kit, Toyobo (Shanghai) Biotechnology Co., Ltd.); according to the sequence information of tomato NHX1 published on NCBI (Access NO.: AJ306630.1; Venema K, Belver A, Marin-Manzano MC, Rodriguez-Rosales MP ,Donaire JP.A novel intracellular K + / H + Antiporter related to Na + / H + Antiporters are important for K + ionhomeostasis in plants.J Biol Chem.2003,278:22453-22459), with Oligo6.0 primer design software (Germany Merck comp...
Embodiment 2
[0044] 1) The cloning of the wild eggplant salt tolerance gene StNHX1 is the same as in Example 1.
[0045] 2) Construction of intermediate vector pCAMBIA3301-121
[0046] pBI121 (Promega, Promega (Beijing) Biotechnology Co., Ltd.) and pCAMBIA3301 (CAMBIA, Australia) were digested with Hind III / EcoR I respectively, and the small fragment (3005bp) in pBI121 and the large fragment (11251bp) in pCAMBIA3301 were recovered ), according to the ratio of 4:1 with T 4 DNA ligase (TaKaRa) ligation, named pCAMBIA3301-121, the ligation product was transformed into DH5α competent cells, and the plasmid was extracted for Hind III / EcoR I double digestion verification. The specific steps are as follows:
[0047] ①Take 10 μL each of pBI121 and pCAMBIA3301 plasmids, and digest them with Hind III / EcoR I respectively. Reaction system for enzyme digestion (50 μL): ddH 2 O 31.5 μL, 10×Buffer E 5 μL, BSA 0.5 μL, pBI121 or pCAMBIA3301 plasma 10 μL, Hind III (Promega) 1.5 μL, EcoR I (Promega) 1.5 μL...
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