Plant low temperature growth associated protein, code genes and application thereof
A gene and protein technology, applied in the field of plant low temperature growth related proteins and their coding genes and applications, can solve the problems of low temperature growth gene isolation and cloning that have not been reported, and achieve the goal of improving low temperature resistance and low temperature tolerance Effect
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Embodiment 1
[0041] Example 1. Discovery of OsCKI1 and its coding genes
[0042] 1. Genetic analysis and preliminary location of genes controlling low temperature growth in rice
[0043] From a set of chromosomal fragment substitution lines carrying IR24 inserts with Asominori as the genetic background (the substitution line has 66 families in total), it is found that one family CSSL13 has a low temperature sensitive phenotype, and this family carries the IR24 insert on the long arm of the second chromosome . The F1 of CSSL13 crossed with the background parent Asominori showed the Asominori phenotype, confirming that the phenotype sensitive to low temperature is a recessive phenotype. By investigating the segregation ratio of 144 F2 plants, we found that 36 plants showed the same phenotype as CSSL13 (3:1 ratio; χ 2 ≈2.68figure 1 a).
[0044] The preliminary positioning method is as follows:
[0045] (1) Extract the total DNA of the selected single plant as a template, the specific method is as f...
Embodiment 2
[0087] Example 2. Obtaining and identification of transgenic plants
[0088] 1. Recombinant expression vector construction
[0089] The pMD18-OsCKI1 constructed in step 3 of Example 1 was digested with HindIII and SacI, and the fragment containing the OsCKI1 gene was recovered and ligated to the pGA1611 vector (uniprotNo: 245199, the network link was http: / / www.uniprot.org / taxonomy / 245199;GENBANKACCESSION NO.AY373338.1), get the recombinant expression vector.
[0090] The recombinant expression vector was transformed into E. coli DH5a competent cells. The plasmid was extracted for restriction enzyme digestion and sequencing. The test results showed that a recombinant plasmid containing the OsCKI1 gene was obtained, which was named pGA1611-OsCKI1 (the backbone vector was pGA1611, and the OsCKI1 shown in sequence 2 was inserted between HindIII and SacI sites. gene).
[0091] 2. Obtaining recombinant Agrobacterium
[0092] The pGA1611-OsCKI1 was transformed into Agrobacterium EHA105 st...
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