Glycosyltransferases play an important role in the synthesis of glycoproteins. Here we report the isolation of a brain cDNA coding for 89% of the porcine UDP-N-acetylglucosamine:alpha-6-D-mannoside-beta-1,2-N-acetylglucosaminy ltransferase II (EC 2.4.1.143) (GnTII). The cDNA was used for screening a genomic liver DNA library and isolation of a recombinant lambda FIX II phage containing the complete porcine GnTII gene and upstream and downstream sequences. The beta-1,2-N-acetylglucosaminyltransferase II gene harbours a single exon with an open reading frame of 1338 bp coding for a 446 amino acid protein with a calculated molecular mass of 51.1 kDa. The promoter of the GnTII gene is lacking a TATA-box and shows variable transcription start sites. In the 3'-untranslated region a polymorphic polyadenosine stretch was detected. The porcine GnTII gene contains four polyadenylation sites. PCR analysis of a porcine-rodent hybrid cell panel revealed the chromosomal location of the GnTII gene on SSC 1q23-q27. The mapping data of the cell panel were confirmed by fluorescence in situ hybridization (FISH) on metaphase chromosomes.
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Biochem Biophys Res Commun
August 2020
International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita-shi, Osaka, 565-0871, Japan. Electronic address:
N-Acetylglucosaminyltransferase II (GNTII), which catalyzes the transfer of N-acetylglucosamine to N-glycans, plays an essential role in the biosynthesis of branched and complex-type N-glycans. Some characteristics of the GNTIIs from various species have been identified, but not all features have been revealed because some insects have GNTII redundancies due to the possession of splicing variants. In this study, we focused on four splicing variants of silkworm Bombyx mori GNTII (BmGNTII) that differ only in the absence or presence of Exon 2, Exon 9 or both, and we characterized the spatiotemporal transcript levels and enzymatic properties of each.
View Article and Find Full Text PDFPLoS One
August 2020
Doctoral Degree Program in Marine Biotechnology, National Sun Yat-sen University, Kaohsiung, Taiwan.
Glycosylation can affect various protein properties such as stability, biological activity, and immunogenicity. To produce human therapeutic proteins, a host that can produce glycoproteins with correct glycan structures is required. Microbial expression systems offer economical, rapid and serum-free production and are more amenable to genetic manipulation.
View Article and Find Full Text PDFJ Biosci Bioeng
March 2019
Laboratory of Biotechnology, Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Laboratory of Biotechnology, Department of Agriculture, Graduate School of Integrated Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan. Electronic address:
N-glycans are involved in various physiological functions and their structures diverge among different phyla and kingdoms. Insect cells mainly produce high mannose-type and paucimannose-type glycans but very few mammalian-like complex-type glycans. However, many insects possess genes for proteins homologous to the enzymes involved in complex-type N-glycan synthesis in mammalian cells, and their N-glycosylation pathway is incompletely understood compared with that of mammals.
View Article and Find Full Text PDFBiotechnol Bioeng
February 2018
Centre for Process Systems Engineering, Department of Chemical Engineering, Imperial College London, London, UK.
Transient gene expression (TGE) is a methodology employed in bioprocessing for the fast provision of recombinant protein material. Mild hypothermia is often introduced to overcome the low yield typically achieved with TGE and improve specific protein productivity. It is therefore of interest to examine the impact of mild hypothermic temperatures on both the yield and quality of transiently expressed proteins and the relationship to changes in cellular processes and metabolism.
View Article and Find Full Text PDFVirus Res
June 2017
International Center for Biotechnology, Osaka University, Yamada-oka 2-1, Suita, Osaka 565-0871, Japan. Electronic address:
Hepatitis B virus (HBV) is the smallest partially double-stranded DNA virus known to infect humans. Worldwide, more than 50% of hepatocellular carcinoma (HCC) cases are related to chronic Hepatitis B. Development of HCC from normal liver cells is characterized by changes in cell surface N-glycans, which can promote the invasive behavior of tumor cells, leading ultimately to the progression of cancer.
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