Journal of Microbiology

Research Support, Non-U.S. Gov't

cDNA Cloning of Korean Human Norovirus and Nucleotidylylation of VPg by Norovirus RNA-Dependent RNA Polymerase: Byung Sup Min , Kang Rok Han , Jung Ihn Lee , Jai Myung Yang; J. Microbiol. 2012;50(4):625-630. Published online August 25, 2012; DOI: https://doi.org/10.1007/s12275-012-2087-4

Abstract: Norovirus, a member of the Caliciviridae family, is a major causative agent of gastroenteritis worldwide. The cDNA of the entire genome of human norovirus (HuNV) was cloned using the RNA extracted from the stool sample of a Korean patient. The RNA genome consists of 7,559 nucleotides, carries 3 open reading frames (ORFs), 5' and 3' noncoding regions, and a poly(A) tail at the 3' end. Phylogenic analysis of the nucleotide sequence indicated that it belongs to GII.4, the most dominant genogroup. To analyze RNA synthesis and nucleotidylylation of VPg by RNA-dependent RNA polymerase (RdRp), recombinant RdRp and VPg were expressed in Escherichia coli as His-tagged forms. The HuNV RdRp exhibited template and divalent cation-dependent RNA synthesis in vitro. The HuNV RdRp nucleotidylylated HuNV VPg but not murine norovirus (MNV) VPg, whereas MNV RdRp nucleotidylylated both MNV and HuNV VPg more efficiently than HuNV RdRp.

Cloning, Expression in Escherichia coli, and Enzymatic Properties of a Lipase from Pseudomonas sp. SW-3: Sun-Young An , Sang-Wan Kim , Yong-Lark Choi , Young-Su Cho , Woo-Hong Joo , oung-Choon Lee; J. Microbiol. 2003;41(2):95-101.

Abstract: The lipase gene (lipA) and its activator gene (lipB) of Pseudomonas sp. SW-3 were cloned and sequenced. The lipB was found to be present immediately downstream of lipA. The deduced amino acid sequences of lipA and lipB showed a high level of homology to those of other lipases belonging to the family I.1 of bacterial lipases. When lipA was expressed in Escherichia coli using T7 promoter, an active lipase was produced in cells carrying both lipA and lipB, but not in cells harboring only lipA. Recombinant lipase (rPSL) overproduced in an insoluble form was solubilized in the presence of 8 M urea, purified in a ureadenatured form and refolded by removing urea in the presence of the Ca^2+ ion. RPLS had maximum activity at pH 8.0 and 50℃, was stable at pHs from 7.0 to 9.0 and below 50 ℃, and showed the highest activity toward the p-nitrophenyl ester of palmitate (C16).

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