Method for promoting plant roots to grow by using BGIos244 genes
A technology of transgenic plants and plant roots, which is applied in the field of promoting the growth of plant roots, can solve the problems of failing to raise a set, and achieve the effects of realizing sustainable agricultural development, promoting plant root growth, and reducing application
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Embodiment 1
[0041] Example 1: PCR amplification of BGIos244 gene and construction of pMD18-T+BGIos244 recombinant vector
[0042] 1. PCR amplification
[0043] The genome of Oryza.rifupongon Yuanjiang (provided by Dong Yang, Kunming Institute of Zoology, Chinese Academy of Sciences) was extracted using a plant genomic DNA extraction kit (TIANGEN New Plant Genomic DNA Extraction Kit, catalog number: DP320-02) DNA (gDNA). According to the base sequence of the BGIos244 gene, a pair of PCR-specific amplification primers were designed: the upstream primer F1 (SEQ ID NO: 1) contained the restriction site BamHI, and the downstream primer R1 (SEQ ID NO: 2) contained the restriction site Restriction site Xba I. Using the Yuanjiang gDNA extracted above as a template, use upstream primer F1, downstream primer R1 and high-fidelity Ex Taq TM (TaKaRa, DRR100B) polymerase for PCR amplification. The PCR amplification system is shown in Table 1.
[0044] Table 1: PCR amplification system of BGIos244 ...
Embodiment 2
[0059] Embodiment 2: Construction of p6 recombinant vector
[0060] 1. PCR amplification of maize ubiquitin (ubi) promoter fragment and construction of pMD18-T+Ubi recombinant vector
[0061] PCR amplification of Ubi promoter
[0062] Genomic DNA (gDNA) of maize variety B73 (Zea mays mayscv.B73) was extracted using a plant genomic DNA extraction kit (TIANGEN New Plant Genomic DNA Extraction Kit, catalog number: DP320-02). According to the method described in Example 1, using the following primers, using the above extracted gDNA of maize B73 as a template, high-fidelity Ex Taq TM (TaKaRa, DRR100B) polymerase for PCR amplification of the Ubi promoter:
[0063] Upstream primer F2 (SEQ ID NO: 3): GG CTGCAG TGCAGCGTGACCCGGTCGT, containing the restriction enzyme site PstI;
[0064] Downstream primer R2 (SEQ ID NO: 4): GG CTGCAG AAGTAACACCAAAC, containing the restriction site PstI.
[0065] The PCR amplified products were separated by 1.0% agarose gel electrophoresis, and t...
Embodiment 3
[0092] Embodiment 3: Construction of p6+BGIos244 recombinant vector
[0093] Using the method described above, the pMD18-T+BGIos244 recombinant vector was extracted, digested with restriction endonuclease BamHI / XbaI, and the BGIos244 gene fragment was recovered. Similarly, the p6 recombinant vector was extracted, digested with the corresponding restriction endonucleases BamHI / XbaI and a large fragment was recovered. Using the method described above, the recovered BGIos244 gene fragment and the p6 recombinant vector fragment were ligated, and the ligated product was transformed into DH5α, thereby finally obtaining the recombinant vector p6+BGIos244. Perform sequencing verification to confirm the correct insertion of the target fragment.
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