A lycopene-producing rhodobacter sphaeroid engineering strain and its construction method
A technology of Rhodobacter sphaeroides and lycopene, which is applied in the field of metabolic engineering, can solve the problems of complex metabolic regulation, increased production cost and safety, and lack of molecular manipulation technology of B. trispora, and achieves the effect of strong strain stability.
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[0063] according to figure 1 Lycopene biosynthetic pathway in the shown Rhodobacter sphaeroides, construction of Rhodobacter sphaeroids engineering strains producing lycopene, the specific construction method is as follows:
[0064] 1. crtI of Rhodospirillum rubrum 4 Gene replacement of crtI in Rhodobacter sphaeroides 3 Gene
[0065] Using the gene of Rhodospira rubrum ATCC 11170 (NCBI website: https: / / www.ncbi.nlm.nih.gov / nuccore / NC_007643.1) as a template, the primer crtI was designed according to the principles of overlap extension PCR primer design 4 -F and crtI 4 -R, using the fidelity enzyme Pfu PCR to amplify the phytoene four-step dehydrogenase gene crtI of Rhodospirillum rubrum ATCC 11170 4 , the PCR amplification system is: 2×Pfu PCR mix 10μL, crtI 4 -F (10 μM) 1 μL, crtI 4 -R (10 μM) 1 μL, Rhodospirillum rubrum gene (20 μg / μL) 1 μL, ddH 2 O7 μL, the reaction program was: denaturation at 94°C for 3 min, followed by denaturation at 94°C for 30 s, annealing at 6...
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