Vegetable oil and fat metabolism related regulatory protein GmMYB73 as well as coding gene and application thereof
A technology for vegetable oils and related proteins, which is applied in the fields of plant genetic improvement, botanical equipment and methods, and applications, and can solve problems such as poorly understood lipid synthesis.
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Embodiment 1
[0066] Example 1. Screening and cDNA Cloning of Transcription Factor GmMYB73 Encoding Gene Related to Regulation of Soybean and Lipid Metabolism
[0067] 1. Fatty acid accumulation and metabolism analysis during soybean seed development
[0068] Two soybean varieties or germplasms with different fatty acid contents: Heinong 44 (HN44) and ZYD7. Among them, the oil content of HN44 grains was 23%, while that of ZYD7 grains was 12%. Such as figure 1 As shown, the seven different stages of the seed development process of the two varieties were collected and preserved separately.
[0069] The oil content of HN44 and ZYD7 seeds in each of the seven developmental stages was measured by gas chromatography, and the oil accumulation curve during seed development was analyzed ( figure 2 ). Such as figure 2 The two inflection points of the oil content growth curve indicated by the arrows were taken respectively: HN44-5 and HN44-6 in HN44; ZYD7-4 and ZYD7-5 in ZYD7. The total RNA of t...
Embodiment 2
[0077] Embodiment 2, the acquisition of GmMYB73 transgenic Arabidopsis
[0078] 1. Construction of plant expression vector pPROKII-GmMYB73
[0079] 1. Primer design
[0080] Primers were designed according to the cDNA sequence (DQ822927) of GmMYB73, and the primer sequences were as follows:
[0081] GmMYB73 up (upstream primer with BamHI restriction site):
[0082] 5'-ggatccAATGGCTGACATAGATC-3';
[0083] GmMYB73dp (downstream primer with KpnI restriction site):
[0084] 5'-ggtACCGATCATTGGCTAGTCG-3'.
[0085] 2. Construction of plant expression vector pPROKII-GmMYB73
[0086] ①Develop Heinong 44 to figure 1 The grains of the 6th stage shown (that is, the 37th day after flowering) were ground in liquid nitrogen, suspended in 4mol / L guanidine thiohydrogen, extracted with acidic phenol and chloroform, and absolute ethanol was added to the supernatant to precipitate the total RNA. cDNA was synthesized from RNA using reverse transcriptase.
[0087] ② Using cDNA as a templat...
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