Cucumber CsDIR16 gene and application of cucumber CsDIR16 gene in lowering plant propamocarb hydrochloride pesticide residues
A technology for pesticide residues and propamocarb, applied in the field of genetic engineering applications, can solve problems such as unreported and reduced propamocarb residues, and achieve the effects of reducing pesticide residues, reducing pesticide usage, and reducing production costs
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Embodiment 1
[0025] CsDIR16 Gene isolation and cloning and expression analysis
[0026] In the previous study, we identified a significantly up-regulated expression induced by propamocarb by analyzing the expression profile of cucumber downy mildew stress. CsDIR16 gene, in order to obtain its full-length sequence information, we searched in the cucumber genome database (http: / / www.icugi.org / ) CsDIR16 The full-length sequence was designed using Primer Premier 5.0 to design specific primers for PCR amplification. The primer sequences are as follows:
[0027] Forward primer: 5′-ATGGCTGGAATCTCTCCAAT-3′
[0028] Reverse primer: 5′-TCAATAATGAAGCACGTAAATGTTA-3′
[0029] Start with Trizol (Invetrigen TM) method to extract cucumber fruit, stem, leaf and total RNA (the kit was purchased from Invetrigen, and the operation method was in accordance with the instructions). Then take 1 ul of the RNA sample, and reverse transcribe it into the first strand of cDNA according to the instructions of the...
Embodiment 2
[0040] CsDIR16 Three-dimensional molecular structure diagram of protein
[0041] Use CPHmodels 3.2 (http: / / www.cbs.dtu.dk / services / CPHmodels / ) to generate atomic coordinates (PDB format), and then use Rasmol windows software to output the three-dimensional molecular structure diagram of CsDIR protein, analyze and predict the protein structure ( Figure 4 ), the protein contains 7 β-sheets (yellow) and 1 α-helix (pink).
Embodiment 3
[0043] CsDIR Family protein and phylogenetic analysis
[0044] Arabidopsis AtDIR The protein is used as the query sequence, and all annotated protein sequences of BLAST cucumber have an e-value value of 1e-20. The results showed that there are a total of 23 in cucumber CsDIR Family genes, sorted according to the distribution of genes on chromosomes, named CsDIR1 , CsDIR2 ... CsDIR23 , as shown in Table 1, 23 CsDIR The genes are distributed on 7 chromosomes of cucumber, among which chromosome 4 is the most. will be 23 CsDIR Compare the proteins of the family genes to construct a gene phylogenetic tree, such as Figure 5 As shown, these CsDIR genes were divided into four subfamilies a, b, d, and e according to the phylogenetic analysis, none of which belonged to the c subfamily. DIRa , DIRb , DIRd with DIRe The differentiation in genetic distance suggests that the possible biological functions of members of this subfamily are not limited to disease resist...
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