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KMID : 0880220200580010039
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Journal of Microbiology
2020 Volume.58 No. 1 p.39 ~ p.45
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Identification and characterization of a novel light-induced promoter for recombinant protein production in Pleurotus ostreatus
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Yin Chaomin
Fan Xiuzhi
Ma Kun
Chen Zheya
Shi Defang
Yao Fen
Gao Hong
Ma Aimin
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Abstract
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A lectin gene (plectin) with a high level of expression was previously identified by comparative transcriptome analysis of Pleurotus ostreatus. In this study, we cloned a 733-bp DNA fragment from the start codon of the plectin gene. Sequence analysis showed that the plectin promoter (Plp) region contained several eukaryotic transcription factor binding motifs, such as the TATA-box, four possible CAAT-box, light respon-siveness motifs and MeJA-responsiveness motifs. To deter-mine whether the Plp promoter was a light-regulated promoter, we constructed an expression vector with the fused egfp-hph fragment under the control of the Plp promoter and transformed P. ostreatus mycelia via Agrobacterium tunte-faciens. PCR and Southern blot analyses confirmed the Plp-egfp-hph fragment was integrated into the chromosomal DNA of transformants. qRT-PCR, egfp visualization, and intracellular egfp determination experiments showed the Plp promoter could be a light-induced promoter that may be suitable for P. ostreatus genetic engineering. This study lays the foundation for gene homologous expression in P. ostreatus.
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KEYWORD
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Pleurotus ostreatus, lectin, promoter, Agrobacterium tumefaciens, light-induction
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