Alpha-tubulin promoter from Chlorella vulgaris allows genetic transformation of green coccoid microalga

作者: Raquel Fernández-Rodríguez , Giovanni Garro-Monge , Maritza Guerrero-Barrantes , Olman Gómez-Espinoza

DOI: 10.18845/TM.V33I2.4155

关键词: Transformation (genetics)Codon usage biasChlorella sorokinianaBiologyGeneticsChlorella vulgarisTATA boxStart codonPromoterCAAT box

摘要: Microalgae have become a feasible platform for high-value recombinant protein production. Although diverse sets of genetic tools microalgae transformation been developed, some critical elements, such as endogenous promoters, need to be improved increasing transgene expression levels after transformation. This work aims evaluate sequence from the 5′ upstream region alpha-tubulin gene Chlorella vulgarisas promoter an antibiotic resistant in sorokiniana. Using silicoanalysis it was possible identify proximal corresponding 281 nucleotides ATG start codon, which possessed 9 potential cis-regulatory element, including TATA Box and CAAT Box. The sequencewas used drive streptomycin-resistance (aada), previous optimization its codon usage. aadasequence allowed obtain GC content 69.8% (compared 53% original sequence), increases similarity with Chlorellasp. genomes (67.2%). Both elements were cloned into pUC57-Kan vector transformed C. sorokiniana cells by electroporation. transgenic colonies identified through culture selective medium PCR. Our results proved capacity vulgaris alpha-tubulin express foreign antibiotic-resistance sorokinianacells. Research promoting sequences is essential order accomplish efficient heterologous expression, especially industrial production like those genus.

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