Construction of a broad host range expression plasmid vector by Golden Gate cloning
Malaysian Journal of Microbiology
; : 588-592, 2021.
Article
em En
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| ID: wpr-973865
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WPRO
ABSTRACT
Aims@#Expression of recombinant proteins across a range of different host organisms has profound contribution to the advancement in biotechnology. In this study, we aimed to construct a highly versatile broad host range (BHR) expression vector, designated as pYL101C.@*Methodology and results@#The Golden Gate cloning approach was used to construct pYL101C. Key features of pYL101C include a strong integron promoter (PINTc), a BHR pBBR1 origin of replication (ori), gentamycin resistance gene (GmR) as a selectable marker and a multiple cloning site (MCS) downstream of the promoter for easy-cloning purpose. To verify the functionality of pYL101C, we cloned the superfolder green fluorescent protein (sfGFP) reporter gene into pYL101C and transferred the resultant recombinant plasmid pYL101C::sfGFP into various Gram-negative bacteria. Transformants obtained stably expressed strong green fluorescence under blue light excitation even without selection after four passages. @*Conclusion, significance and impact of study@#The constructed BHR expression vector, pYL101C and recombinant pYL101C::sfGFP are stable and can be used to monitor the presence of Gram-negative bacteria, such as endophytes and pathogens in their hosts and environment.
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Base de dados:
WPRIM
Assunto principal:
Plasmídeos
/
Clonagem Molecular
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Especificidade de Hospedeiro
Idioma:
En
Revista:
Malaysian Journal of Microbiology
Ano de publicação:
2021
Tipo de documento:
Article