Plant flowering regulation related protein TaSOD as well as coding gene and application thereof
A technology related to proteins and plants, applied in the field of genetic engineering
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Embodiment 1
[0032] Embodiment 1: Cloning and sequence motif analysis of TaSOD gene
[0033] The leaves of wheat photothermosensitive male sterile line BS366 were taken, and the total RNA of anthers was extracted by Trizol method. cDNA was obtained by reverse transcription with superscript II (purchased from Invitrogen) reverse transcriptase. Primers P1 and P2 were designed according to the sequence of TaSOD gene coding region. Using the cDNA obtained by reverse transcription as a template, PCR amplification was performed with primers P1 and P2. The sequences of primers P1 and P2 are as follows:
[0034] P1: 5'-ATGGCAGGGAAACCCGGCA-3'
[0035] P2: 5'-TCAAAATGTATGATCTTCAACGATATC-3'.
[0036] The PCR product was detected by 1% agarose gel electrophoresis, and a band with a molecular weight of about 600bp was obtained, which was consistent with the expected result. This fragment was recovered using an agarose gel recovery kit (TIANGEN). The recovered fragment was ligated with pGEM-TEasy ...
Embodiment 2
[0038]Embodiment 2: Cultivate early flowering transgenic plant with TaSOD gene
[0039] 1. Construction of recombinant expression vector
[0040] Construction of pBI29A-TaSOD Dicotyledon Recombinant Expression Vector
[0041] Using the cDNA obtained by reverse transcription of the total RNA of the leaves of the wheat photothermosensitive male sterile line BS366 as a template, PCR amplification was performed with specific primers containing SacI and SpeI linker sequences; then SacI and SpeI double-digested the PCR product, recovered, The digested product was inserted into vector pBI121 between the SacI and SpeI restriction sites behind the CaMV 29A promoter, to obtain the recombinant vector pBI29A-TaSOD.
[0042] The primer sequences are as follows:
[0043] TaSOD[SacI]: 5'–TCCGAGCTCTCAATCAATGTCAGCGTGCAC-3'
[0044] TaSOD[SpeI]: 5'– CGGACTAGT ATG CCG GTG CTG GCG ATG-3'.
[0045] 2. Acquisition and identification of transgenic Arabidopsis
[0046] Obtaining transgenic ...
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