Protein related to plant epidermal hair growth, coding gene of protein related to plant epidermal hair growth and application
A technology encoding genes and proteins, applied in the direction of plant gene improvement, application, plant peptides, etc., to achieve the effect of inhibiting the formation of epidermal hair
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Embodiment 1
[0036] Embodiment 1, the cloning of the gene CsTRY related to the development of plant epidermis
[0037] 1. Materials
[0038]Cucumber Jinyan No. 4 (purchased from the Cucumber Research Institute of Tianjin Academy of Agricultural Sciences) was used as the test material. This variety has obvious hairy skin and dense thorn tumors. Cucumber seeds were sown in a solar greenhouse for ordinary cultivation and management. When the plants began to produce melons, the fruits on the day of flowering were taken and frozen in liquid nitrogen, and stored in a -80°C refrigerator for later use.
[0039] 2. Total RNA extraction and cDNA synthesis
[0040] 1. Extraction of total RNA
[0041] The total RNA of cucumber fruit was extracted with a column plant RNAout kit (purchased from Beijing Tianenze Gene Technology Co., Ltd., catalog number 71203), and the residual DNA in the RNA sample was removed with DNase. Proceed as follows:
[0042] (1) Add about 200 mg of cucumber fruit into a mor...
Embodiment 2
[0075] Embodiment 2, the acquisition of transgenic plants and its detection
[0076] 1. Construction of a recombinant expression vector containing the target gene CsTRY
[0077] 1. Construction of CsTRY overexpression vector
[0078] The positive bacterial plasmid CsTRY-PBSK with correct sequencing and the overexpression vector ms1300 (the public can obtain from China Agricultural University, the non-patent literature that has recorded this material is: Liao Hui. Phd cold resistance research and screening analysis of ABA sensitive mutation. Master Dissertation.p23) Carry out sacI and HindIII double enzyme digestion at the same time, and digest overnight at 37°C. After digestion, the 249bp target fragment and the large fragment of the overexpression vector were recovered, ligated with T4DNA Ligase, and ligated overnight at 16°C to obtain the target plasmid. The target plasmid was transformed into Escherichia coli, resistance screening, positive clones were picked, the positiv...
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