CpSOC1 gene of chimonanthus praecox, protein encoded by CpSOC1 gene and application
A technology of protein and wintersweet, applied in the field of genetic engineering, to achieve the effects of prolonging the persistence time, regulating the flowering period and improving the ornamental value
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Embodiment 1
[0028] Example 1 Cloning and expression analysis of Wintersweet CpSOC1 gene
[0029] 1 Cloning of the CpSOC1 gene of Wintersweet
[0030] 1.1 Extraction of total RNA
[0031] (1) Flower buds and flower buds of Chimonanthus praecox (planted next to the fourth sports ground on the campus of Southwest University, No. 2 Tiansheng Road, Beibei District, Chongqing, routine maintenance and management) (April, May, July, September), Bloom, decay and leaf mix samples. The glassware, mortar, pestle, scissors, tweezers, medicine spoon, etc. used for RNA extraction were soaked in 0.1% DEPC aqueous solution at 37°C for 12h, then wrapped in tin foil and sterilized by dry heat in an oven at 180°C for 4h, then cooled spare.
[0032] The RNA was extracted using the RNAprep Pure Plant Total RNA Extraction Kit produced by Beijing Tiangen Company. The extraction process was slightly modified according to the instructions. The specific operations are as follows:
[0033] (1) Transfer 500 μL of...
Embodiment 2
[0116] Example 2 Agrobacterium-mediated transformation of Wintersweet CpSOC1 gene (Arabidopsis thaliana)
[0117] 1 Construction of expression vector
[0118] According to the requirements of the experiment, in this embodiment, the characteristic of the multiple cloning restriction site on the vector pCAMBIA2301g was utilized, and the cDNA fragment on the cloning vector was digested and ligated into the expression vector.
[0119] 2. Preparation of Agrobacterium GV3101 and EHA105 Electrically Competent Cells and Transformation of Expression Vectors
[0120] (1) Pick fresh single colonies of Agrobacterium GV3101 and EHA105 in 10 mL of LB liquid medium, and cultivate them with shaking at 28°C until the late logarithmic period;
[0121] (2) Take 1 mL of bacterial liquid and add it to 100 mL of fresh LB liquid medium, shake and culture at 28°C until OD 600 About 0.5-1.0;
[0122] (3) Transfer the bacterial solution to a 50mL centrifuge tube and place in an ice bath for 30 minut...
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