Applications of SlSNAT1 protein and related biological materials thereof in regulating aging resistance of plant seeds
A technology of plant seeds and biomaterials, applied in the application field of regulating the aging resistance of plant seeds, to achieve the effects of improving aging resistance, wide application prospects, and increasing expression
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Embodiment 1
[0063] Cloning of Example 1 SlSNAT1 Gene
[0064] 1. Acquisition of experimental materials
[0065] The tomato Micro Tom was planted in the light incubator of the College of Horticulture, China Agricultural University. The light incubator culture conditions were light density 10000LUX; light 12h, temperature 26°C; darkness 12h, temperature 16°C. Tomato seedlings grew to the fourth leaf, and the mature leaves were taken, frozen in liquid nitrogen quickly, and stored in a -80°C refrigerator for later use.
[0066] 2. Extraction of RNA
[0067] RNA extraction kit (purchased from Huayueyang Biotechnology Co., Ltd., catalog number: 0416-50) was used to extract total RNA from tomato leaves.
[0068] 3. Acquisition of cDNA
[0069] Using the total RNA extracted in step 2 as a template, use a reverse transcription kit (purchased from Takara, Cat. No.: RR047A) to perform a reverse transcription reaction to obtain cDNA, and further dilute the cDNA solution to 100 ng / μl as the followi...
Embodiment 2
[0083] Example 2 Application of SlSNAT1 in Germination Regulation of Aged Tomato Seeds
[0084] (1) Construction of SlSNAT1 overexpression vector
[0085] Use the ClonExpress II Recombination Cloning Kit (purchased from Nanjing Novizan Biotechnology Co., Ltd., Cat. No. C112-01) to connect the target gene fragment with the vector to obtain the ligation product; the recombination reaction steps are as follows:
[0086] 1. Insert amplification: By introducing a homologous sequence at the end of the linearized cloning vector at the 5' end of the primer, the 5' and 3' ends of the inserted fragment amplification product are completely consistent with the two ends of the linearized cloning vector. the sequence of.
[0087] According to the above principles and the sequence of the gene SlSNAT1, the amplification primers with the restriction site SmaI and the homologous sequence at the end of the linearized cloning vector were designed at the 5′ end. The primer sequences are as follow...
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