Host microorganisms
A technology of microorganisms and recombinant microorganisms, applied in the field of host microorganisms, can solve problems such as insufficient improvement in productivity
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Embodiment 1
[0037] Genomic DNA was extracted from Bacillus subtilis 168. Using the extracted gene as a template, make a 1.5-kb DNA fragment (A) adjacent to the upstream end of the sigF gene (base number 2442658←2443425) on the genome, and a length adjacent to the downstream end of the sigF gene Multiplied for the 1.5-kb DNA fragment (B). Using plasmid pC194 as a template, a 0.9-kb DNA fragment containing the chloramphenicol resistance gene was individually multiplied (C). Fragments (A), (B), and (C) were sequentially ligated by SOE-PCR to prepare a 3.9-kb DNA fragment. The obtained DNA fragment was used to transform Bacillus subtilis 168 by an appropriate method. Bacillus subtilis 168 was cultured on LB agar medium containing chloramphenicol, and colonies were isolated as transformants. It was confirmed by PCR and sequencing that in the genome of the transformant, the region containing the sigF gene (2442632-2443318) was deleted and replaced by the chloramphenicol-resistant gene. Each...
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