A kind of plant fertility-related protein and its application
A plant and protein technology, applied in the field of plant fertility-related proteins and their applications, can solve the problems of limiting the mining and utilization of soybean germplasm resources, limiting the aggregation and utilization of excellent gene loci, etc.
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Embodiment 1
[0064] Embodiment 1, the acquisition of soybean male sterility gene GmMS1
[0065] The soybean genome was sequenced and functionally analyzed, and a soybean male sterility gene was found, which was named GmMS1 gene. Its CDS is shown in sequence 2 of the sequence listing, and its genome sequence is shown in sequence 7 of the sequence listing. The protein encoded by the GmMS1 gene is named GmMS1 protein, as shown in sequence 1 of the sequence listing.
Embodiment 2
[0066] Example 2, Application of GmMS1 Gene in Regulating Soybean Fertility
[0067] 1. Construction of Crisper / CAS9 gene editing vector
[0068] 1. Design and synthesis of gRNA targets
[0069] Design two target sequences, respectively
[0070] Target 1: GACGGGAACACCTGTGGCGG TGG ;
[0071] Target 2: CGCCGAGGTCTAAGATACAG AGG .
[0072] In target 1 and target 2, the underline is the PAM sequence.
[0073] Two pairs of primers were designed according to the target sequence:
[0074] Target 1 primer:
[0075] F1:5'-TTGGACGGGAACACCTGTGGCGG-3';
[0076] R1: 5'-AACCCGCCACAGGTGTTCCCGTC-3'.
[0077] Target 2 primers:
[0078] F2: 5'-TTGCGCCGAGGTCTAAGATACAG-3';
[0079] R2: 5'-AACCTGTATCTTAGACCTCGGCG-3'.
[0080] 2. Formation of primer dimers
[0081] Dilute primer F1 and primer R1 to 10 μM respectively, and configure the reaction system: F15 μl, R15 μl, H 2 O 15 μl. After mixing, react at 95°C for 3 minutes, then naturally cool to 25°C, and then 16°C for 5 minutes to ...
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