Using green fluorescent protein fusion approach to demonstrate the putative laccase-encoding gene expression and an importance of 3′ untranslated region in gene expression in Penicillium marneffei

Abstract

Penicillium marneffei is an opportunistic fungus that can cause a fatal disseminated infection in immunocompromised patients. Understanding the biology and pathogenic capacity of this fungus is essential for medical treatment and infection control. Genetic transformation has been developed to investigate the expression and function of the gene of interest in pathogenic fungi including P. marneffei. To date, the whole genome sequence of P. marneffei has been submitted to an open access GenBank database enhancing the capability of genetic analysis. The genetic analysis provides an insight in its biology and pathogenicity and an option of a rational target for drug development. In this study, we fused the enhanced green fluorescent protein (egfp) gene into a putative laccase-encoding gene (arb2) of P. marneffei to demonstrate gene expression and the effect of 3′ untranslated region (UTR) to the production of protein. Our findings highlight the necessary of keeping a native promoter and 3′ UTR for gene expression analysis. Many considerations are described and demonstrated for successful fusion protein production. These data should be beneficial for further studies including gene expression, protein localization, and the role of gene and nucleotide sequences.