pheS ^AG Based Rapid and Efficient Markerless Mutagenesis in Methylotuvimicrobium.

ABSTRACT: Due to their fast growth rate and robustness, some haloalkalitolerant methanotrophs from the genus Methylotuvimicrobium have recently become not only promising biocatalysts for methane conversion but also favorable materials for obtaining fundamental knowledge on methanotrophs. Here, to realize unmarked genome modification in Methylotuvimicrobium bacteria, a counterselectable marker (CSM) was developed based on pheS, which encodes the α-subunit of phenylalanyl-tRNA synthetase. Two-point mutations (T252A and A306G) were introduced into PheS in Methylotuvimicrobium buryatense 5GB1C, generating PheS ^AG , which can recognize p-chloro-phenylalanine (p-Cl-Phe) as a substrate. Theoretically, the expression of PheS ^AG in

SUBMITTER: Liu Y 

PROVIDER: S-EPMC7136838 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature