A recombinant vector for assisting hydrogen production, a recombinant strain and its construction method and application
A technology of recombinant strains and recombinant vectors, which is applied in the field of genetic engineering, can solve problems such as differences in hydrogen production efficiency, low hydrogen gas, and inability to meet human needs for clean energy, and achieve the effect of improving hydrogen production efficiency
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[0047] (1) Search for the homologous gene sequence of the Bacillus cereus FHL system transcription protein FhlA on NCBI, and then combine the pET32a plasmid sequence, and use DNAMAN software to design the FHL system transcription protein FhlA encoding gene with the vector homologous sequence primers. The designed primers were handed over to Sangon Bioengineering (Shanghai) Co., Ltd. for primer synthesis.
[0048] The designed primers are:
[0049] Forward-Primer:
[0050] AACTTTAAGAAGGAGATATA CATATG TTTCGAGATAATCGTACAAAT
[0051] Reverse-Primer:
[0052] CAGCCGGATCTCAGTGGTGGTGGTGGTGGTG CTCGAG TAATGGTACATTTACTTTTAC
[0053] (2) According to the designed primers, the fhlA gene fragment was amplified by PCR technique. PCR amplification program: denaturation at 94°C for 15s, annealing at 60°C for 10s, extension at 68°C for 2min, 30 cycles; PCR amplification system: 25μL 2×HiFi-PCRMaster, 20μL ddH 2 O, 1 μL DNA template, 2 μL upstream primer (10 μmol / L), 2 μL downstream p...
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