Salt-tolerant synechococcus genetically engineered bacterium capable of biosynthesizing glycerol glucoside and construction method thereof
A technology of genetically engineered bacteria and glucoside, which is applied in the field of biosynthetic glycerol glucoside and salt-tolerant Synechococcus genetically engineered bacteria and construction, which can solve the problems of limiting large-scale cultivation of cyanobacteria
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Embodiment 1
[0041] 1. Construct the vector pSI-ggpP:
[0042] Use the nucleotide sequences shown in SEQ ID NO.7 and SEQ ID NO.8 as upstream and downstream primers, and use the plasmid pSI-SPE as a template to perform reverse PCR amplification to obtain the pSI-SPE plasmid backbone (SEQ ID NO.1) , using the bacterial genome extraction kit to extract the Synechocystis sp. PCC 6803 genome as a template, and using the nucleotide sequences shown in SEQ ID NO.9 and SEQ ID NO.10 as upstream and downstream primers to perform PCR The gene ggpP (SEQ ID NO.2) encoding glycerol glucoside-phosphate phosphatase was amplified, and the pSI-SPE plasmid backbone was connected with the gene ggpP encoding glycerol glucoside-phosphate phosphatase to form the vector pSI-ggpP;
[0043] 2. Construct the vector pSI-ggpS:
[0044] The bacterial genome extraction kit was used to extract the genome of Synechocystis sp. PCC 6803 as a template, and the nucleotide sequences shown in SEQ ID NO.11 and SEQ ID NO.12 were ...
Embodiment 2
[0048] 1. Construction of vectors pSII-2522 and pSI-2522ggpPS
[0049] (1) Construction of vector pSII-2522: use the nucleotide sequences shown in SEQ ID NO.7 and SEQ ID NO.16 as upstream and downstream primers, and use the plasmid pSII-CM as a template to perform inverse PCR amplification to obtain pSII-CM Plasmid backbone (SEQ ID NO.4); use the bacterial genome extraction kit to extract the Synechococcus Syn2973 genome as a template, and use the nucleotide sequences shown in SEQ ID NO.14 and SEQ ID NO.15 as upstream and downstream primers for PCR The gene Syn7942-2522 (SEQ ID NO.5) encoding glycerol-3-phosphate dehydrogenase was amplified, and the pSII-CM plasmid backbone was ligated with the gene Syn7942-2522 encoding glycerol-3-phosphate dehydrogenase into vector pSII-2522;
[0050] (2) Construction of vector pSI-2522ggpPS:
[0051] Use the nucleotide sequences shown in SEQ ID NO.7 and SEQ ID NO.12 as upstream and downstream primers, and use the plasmid pSI-ggpPS as a te...
Embodiment 3
[0053] Construction of down-regulated sucrose synthesis vector pSII-drfbA:
[0054] Using SEQ ID NO.7 and SEQ ID NO.18 as upstream and downstream primers, and using plasmid pSII-CM as a template, carry out reverse PCR amplification to obtain the pSII-CM plasmid backbone and gene synthesis sequence PpsbA2M-asrfbA-micC-TrbcL-P cpc560 -hfq (SEQ ID NO.6) was ligated and assembled into vector pSII-drfbA (see figure 2 ).
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