Cymbidium goeringii CgWRKY40 gene and application thereof
A gene and amino acid technology, applied in the Chunlan CgWRKY40 gene and its application fields, can solve the problems of affecting the regulation mechanism, gene inactivation, activity enhancement, etc., and achieve the effects of promoting vegetative growth, delaying flowering, and vigorous vegetative growth
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Embodiment 1
[0023] The material used in this example is the leaves of Chunlan 'Songmei', which are quickly frozen in liquid nitrogen after harvest and stored in an ultra-low temperature refrigerator (-80°C).
[0024] 1) Extraction of total RNA from Chunlan leaves
[0025] According to the instructions of the TaKaRa plant total RNA extraction kit, the specific operation is as follows:
[0026] The ultra-low temperature frozen leaves of Chunlan were quickly transferred to a mortar pre-cooled with liquid nitrogen, and the tissue was ground with a pestle, during which liquid nitrogen was continuously added until it was ground into powder; the powdered sample was added to a 450 μl In a 1.5 mL sterilized tube of Buffer PE, pipette repeatedly until there is no obvious precipitation in the lysate; centrifuge the lysate at 12,000 rpm at 4°C for 5 min; carefully pipette the supernatant into a new 1.5 mL sterilized tube . Add 1 / 10 volume of Buffer NB to the supernatant, shake and mix with Vortex, ...
Embodiment 2
[0048] Research indicates WRKY40 The gene was expressed in every tissue and organ of Chunlan ( figure 2 a), but the gene has the highest expression level in spring orchid, indicating that the gene is functionally active in flowers. Through the expression analysis of the leaves of Chunlan ABA stress treatment ( figure 2 b), to prove CgWRKY40 Genes play an important regulatory role in the ABA stress response of Chunlan leaves.
[0049] The plant material used in this embodiment is Arabidopsis ( Arabidopsis thaliana ) Col (Columbia) wild-type seeds.
[0050] The Escherichia coli strain used in this example was Trans5α; the Agrobacterium strain was GV3101, which were used to transform Arabidopsis thaliana; the plant expression vector used in the experiment was pBI121. The strains used were purchased from Quanshijin Biotechnology Company and Price Biotechnology Company.
[0051] 1) CgWRKY40 Construction of gene overexpression vector
[0052] With embodiment 1 obtaining ...
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