Construction of the recombinant nuclear localization expression system in Coprinopsis cinerea [灰盖拟鬼伞核定位蛋白重组表达系统的构建]

作者：Wang Y.; Zhao J.; Tian X.; Jin S.; Ran Y.; Huang X.; Yuan J.; Yuan S.; Liu Z.^*

来源：Mycosystema, 2022, 41(1): 68-77.

DOI：10.13346/j.mycosystema.210189

• green fluorescent protein
• nuclear localization sequence
• promoter
• recombinant expression

To construct a recombinant nuclear localization (NL) expression system for Coprinopsis cinerea, the NL sequence (NLS) of histone H2B was determined by amino acid sequence alignment and informatics analysis. The recombinant expression vector of fusion green fluorescent protein (GFP) with the NLS of histone H2B was constructed and transferred into C. cinerea AmutBmut strain to examine whether this NLS can guide GFP expression to the nucleus. The results showed that the codon-optimized GFP gene was expressed to produce green fluorescence under the control of the β-tubulin promoter of C. cinerea. Furthermore, NLS from histone H2Bb (CC1G_07639) can successfully induce GFP expression in the nucleus. In this study, a recombinant NL expression system for C. cinerea with proven efficiency was constructed. Our findings are beneficial to optimizing the CRISPR/Cas9 gene editing system and further exploring the mechanisms of fruiting body formation in C. cinerea.

• 单位
  南京师范大学