CpSOC1 gene of chimonanthus praecox, protein encoded by CpSOC1 gene and application
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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
Active Publication Date: 2017-11-03
SOUTHWEST UNIV
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However, there is no related report in Chimera chinensis, so this experiment studies the function of Wintersweet CpSOC1 on the basis of previous studies.
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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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Abstract
The invention provides a CpSOC1 gene of chimonanthus praecox, protein encoded by the CpSOC1 gene and application of the gene in regulation of a flowering stage. The chimonanthus praecox CpSOC1 protein provided by the invention is 1) a protein consisting of amino acids shown as SEQ ID NO. 2; or 2) a protein derived from the protein 1) by substitution, deletion or addition of one or more amino acids in an amino acid sequence shown as SEQ ID NO. 1 and having the same activity. The invention also provides a gene for encoding the protein. According to the invention, cloning is carried out to obtain CpSOC1 gene of chimonanthus praecox for the first time, and the function of the gene is verified in a plant (arabidopsis thaliana) by a transgenic technology. Results show that the flowering stage of transgenic arabidopsis thaliana is earlier than that of a control group, and the survival time of calyxes of transgenic Arabidopsis thaliana having a high expression level and a moderate expression level is significantly longer than that of the control group, thereby providing the possibility for the regulation of the flowering stage and the increase of the ornamental value of the plant.
Description
technical field [0001] The invention relates to the field of genetic engineering, in particular to the wintersweet CpSOC1 gene and its application in flowering regulation. Background technique [0002] Chimonanthus praecox (L.) Link, belonging to Chimonanthus praecox (L.) Link, belongs to Chimonanthus Lindl. in the family Calycanthaceae, and is a rare and precious ornamental flower and tree endemic to my country. Wintersweet is a rare winter flowering plant. It flowers first and then leaves. It has sweet flowers, golden color, waxy petals and strong fragrance. It is the best cut flower material in winter. Wintersweet has been widely used in ornamental gardens, and its medicinal and edible value has also been developed and utilized. Wintersweet is a bushy shrub that can reach a height of 3 to 4 meters. Deciduous, opposite leaves, semi-leathery; flowers are born in leaf axils; the outermost perianth is brown and scale-like, and the inner transition is yellow; flowering perio...
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