Tomato gene SLMBP21 and application thereof
A tomato and gene technology, applied in the field of genetic engineering, can solve problems such as unknown functions and different functions, and achieve the effect of good application value
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Embodiment 1
[0030] Example 1 Cloning of full-length tomato SLMBP21 gene
[0031] 1) Using a forward primer with a nucleotide sequence as shown in SEQ ID No.3 and a reverse primer with a nucleotide sequence as shown in SEQ ID No.4, clone SLMBP21 from the pedicel cDNA of tomato Zhongshu No. 4 The sequence of the coding region of the gene;
[0032] PCR program: 94°C, 5 minutes; 94°C, 30 seconds; 55°C, 30 seconds; 72°C, 45 seconds; repeat 35 times; 72°C, 10 minutes.
[0033] PCR system: 2×EasyTaq PCR SuperMix (Quanshijin Company) 25μl;
[0034] Forward primer (10μM) 2μl;
[0035] Reverse primer (10μM) 2μl;
[0036] DNA template 5μl;
[0037] Double distilled water to make up 50μl.
[0038]The above PCR product was directly cloned and connected to pEASY-T1Simple according to the TA cloning method (such as image 3 shown); the ligation product was transformed into Escherichia coli DH5α and multiplied in it, and the positive clone was sequenced and screened to obtain the sequence;
[0039...
Embodiment 2
[0057] Example 2 Sequence Structure Analysis of Tomato SLMBP21 Protein
[0058] Tomato SLMBP21 has the characteristic sequence region of MADS-box family proteins: MADS region (about the first 60 amino acids), which has 69% similarity with SEP4 protein sequence in Arabidopsis and 84% similarity with LeMADS1 protein sequence in tomato . The result is as figure 1 shown.
Embodiment 3
[0059] Example 3: Sequence structure analysis of tomato SLMBP21 protein
[0060] The results of phylogenetic analysis showed that SLMBP21 belonged to the SEP subfamily, and belonged to the same branch of the evolutionary tree as SEP1 / 2 / 3 / 4 in Arabidopsis and Ph FBP2 / 4 / 5 / 9 / 23 in Petunia. Other members of this subfamily in tomato include TM5, TM29, RIN and LeMADS1.
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