Keywords

Spirulina, Plasmid Library, Stk, Cloning

Abstract

Use the improved SDS method to extract the genomic DNA of Spirulina platensis, digest it with Sau3AI, recover 1 to 2 kb fragments, connect them to the pBR322 plasmid vector that has been digested and dephosphorylated with BamHI (the same tail enzyme of Sau3AI), and then transfected. into Escherichiacoli Top10 cells to construct a Spirulina genome plasmid library. Design anchor primers based on the sequences on both sides of the multiple cloning site on the pBR322 vector, design degenerate primers based on the conserved sequence of filamentous cyanobacteria serine/threonine kinase (STK), and use the Spirulina plasmid library as a template for “nested” amplification The partial sequence of the STK gene was published. The construction of Spirulina genome plasmid library provides great convenience for functional gene research.

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Original research done by Tian Li, Zhang Xiaowen, Qin Song

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