Application of E-like gene in rice in cultivating early-flowering and dwarf transgenic plants
A technology of transgenic plants and genes, which is applied in the field of application of rice E genes in the cultivation of early flowering and dwarf transgenic plants, and can solve problems such as weak research
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Embodiment 1
[0056] Example 1, Regulatory function of rice class E genes in flower organ development
[0057] 1. Gene silencing
[0059] The OsMADS8 gene was cloned by the RACE method, and the OsMADS7 gene was obtained by screening the rice cDNA library with the OsMADS8 gene as a probe.
[0060] 1) Construction of rice young panicle cDNA library
[0061] Total RNA was extracted from 1-5 cm young panicles of wild-type rice Zhonghua 11 (purchased from the Institute of Crop Science, Chinese Academy of Agricultural Sciences). After mRNA was isolated, a cDNA library was constructed using Stratagene's ZAP-cDNA Synthesis Kit. After titer detection: the titer of the original library was 3.7×10 6 The number of plaques / ml, the titer of the amplified library was 9.3×10 9 Number of plaques / ml. The identification of blue and white spots shows that the plaque rate is only 0.8%. 17 phage plaques were randomly selected, and plasmids were extracted after circularization, and ...
Embodiment 2
[0145] Embodiment 2, the overexpression of gene
[0146] 1. Gene overexpression
[0147] 1. Construction of overexpression vector
[0148] In order to facilitate the detection of protein levels in transgenic plants, we fused the MYC-FLAG tag in front of or behind the OsMADS8 coding region; the Ubiqutin promoter is a promoter with strong expression in monocotyledonous plants, and the Ubiqutin promoter used in Example 2 The promoter is the promoter prepared in Example 1. We chose this promoter to construct two OsMADS8 overexpression vectors: pU1301MFOs8 and pU1301Os8MPF ( Figure 24 ).
[0149] Primers H8F and H8R were designed according to the OsMADS8 sequence, and the forward primer H8F was introduced into the BamHI site in front of the start codon. The sequence is: 5'ACTA ggATCC ATggggAgAgggAgggT-3'; H8R reverse primer, mutating the stop codon TGA in the OsMADS8 sequence to ATC (Ile) and introducing a ClaI site, the sequence is: 5'-CAgg ATGAT gggTAgCCATgTCggCA-3'. The...
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