• Molecules and Cells
• /
• v.44 no.1
• /
• pp.26-37
• /
• 2021

Human papillomaviruses (HPVs) cause cellular hyperproliferation-associated abnormalities including cervical cancer. The HPV genome encodes two major viral oncoproteins, E6 and E7, which recruit various host proteins by direct interaction for proteasomal degradation. Recently, we reported the structure of HPV18 E7 conserved region 3 (CR3) bound to the protein tyrosine phosphatase (PTP) domain of PTPN14, a well-defined tumor suppressor, and found that this intermolecular interaction plays a key role in E7-driven transformation and tumorigenesis. In this study, we carried out a molecular analysis of the interaction between CR3 of HPV18 E7 and the PTP domain of PTPN21, a PTP protein that shares high sequence homology with PTPN14 but is putatively oncogenic rather than tumor-suppressive. Through the combined use of biochemical tools, we verified that HPV18 E7 and PTPN21 form a 2:2 complex, with a dissociation constant of 5 nM and a nearly identical binding manner with the HPV18 E7 and PTPN14 complex. Nevertheless, despite the structural similarities, the biological consequences of the E7 interaction were found to differ between the two PTP proteins. Unlike PTPN14, PTPN21 did not appear to be subjected to proteasomal degradation in HPV18-positive HeLa cervical cancer cells. Moreover, knockdown of PTPN21 led to retardation of the migration/invasion of HeLa cells and HPV18 E7-expressing HaCaT keratinocytes, which reflects its protumor activity. In conclusion, the associations of the viral oncoprotein E7 with PTPN14 and PTPN21 are similar at the molecular level but play different physiological roles.

• Parasites, Hosts and Diseases
• /
• v.59 no.1
• /
• pp.35-45
• /
• 2021

Adult echinostomes having 37 collar spines collected from the intestine of Pitalah ducks in Aceh Province, Indonesia in 2018 were morphologically and molecularly determined to be Echinostoma miyagawai Ishii, 1932 (Digenea: Echinostomatidae). Among 20 ducks examined, 7 (35.0%) were found to be infected with this echinostome, and the number of flukes collected was 48 in total with average 6.9 (1-17) worms per duck. The adult flukes were 7.2 (6.1-8.5) mm in length and 1.2 (1.0-1.4) mm in width (pre-ovarian or testicular level) and characterized by having a head collar armed with 37 collar spines (dorsal spines arranged in 2 alternating rows), including 5 end group spines, and variable morphology of the testes, irregularly or deeply lobed (3-5 lobes) at times with horizontal extension. The eggs within the worm uterus were 93 (79-105) ㎛ long and 62 (56-70) ㎛ wide. These morphological features were consistent with both E. miyagawai and Echinostoma robustum, for which synonymy to each other has been raised. Sequencing of 2 mitochondrial genes, cox1 and nad1, revealed high homology with E. miyagawai (98.6-100% for cox1 and 99.0-99.8% for nad1) and also with E. robustum (99.3-99.8% for nad1) deposited in GenBank. We accepted the synonymy between the 2 species and diagnosed our flukes as E. miyagawai (syn. E. robustum) with redescription of its morphology. Further studies are required to determine the biological characteristics of E. miyagawai in Aceh Province, Indonesia, including the intermediate host and larval stage information.

• Journal of Food Hygiene and Safety
• /
• v.35 no.6
• /
• pp.574-580
• /
• 2020

In this study we sought to determine the food fraud by discriminating species of commercial seafood product such as Larimichthys polyactis, Larimichthys crocea, Pennahia argentatus, and Miichthys miiuy, which are difficult to morphologically discriminate. After amplifying the mitochondrial cytochrome c oxidase subunit I gene of the reference fish, the DNA sequences of the amplified PCR products were analyzed. As a result, a 655 bp sequence for species identification was selected for use as DNA barcodes. To confirm the DNA data and primer set, the DNA barcode sequence of each fish was compared to that in that in the NCBI. All of the DNA barcode data were matched with the gene sequence of each fish in the NCBI. A total of 32 processed seafood products (8 L. polyactis, 12 L. crocea, 3 Pennahia argentatus, and 9 Miichthys miiuy) were investigated. Homology of 97% or more in DNA sequences was judged as the same species. As a result of the monitoring, there were no discovered cases of forgery or alteration. However, the use of a raw material name having no matching standard name in the Korea Food Code may cause consumer confusion. Therefore, it is suggested that the standard name or scientific name be co-labeled with the raw material name on seafood products to prevent consumer confusion.

• Journal of the Korean Applied Science and Technology
• /
• v.39 no.5
• /
• pp.605-613
• /
• 2022

The genus Flavobacterium, type genus of the family Flavobacteriaceae and a member of the phylum Bacteriodetes includes gram-negative and yellow-pigmented rods. Those bacteria have been isolated from various environments of the earth. A yellow-pigmented, gram-negative rod was isolated from a pond in the campus of the Changwon University, Changwon, Kyeongnam and designated as strain B1. Strain B1 was further analyzed physiologically, biochemically and phylogenetically, and concluded to be a member of genus Flavobacterium. BLAST search of the 16S rRNA gene sequence of strain B1 shows homology no higher than 99.0% with those sequences of other bacteria. The major fatty acids of strain B1 are iso-C_15:0 (19.6%), summed feature 3(C_16:1 ω7c and/or C_16:1 ω6c, 16.1%), iso-C_17:0 3OH(10.2%), iso-C_15:0 3OH(8.4%) and iso-C_15:1 G(6.6%) showing significant differences in fatty acid compositions between strain B1 and the other known Flavobacterium species. DNA sequence of 16S rRNA gene of strain B1 was deposited in genbank under accession number OP060681.

• Journal of Life Science
• /
• v.31 no.7
• /
• pp.671-676
• /
• 2021

A cDNA encoding the protein lethal (2) essential for life [l(2)efl] was cloned from Gryllus bimaculatus and named GBl(2)efl. This protein is composed of 189 amino acids, including an N-glycosylation site and 15 phosphorylation sites. Its predicted molecular mass is 21.19 kDa, with a theoretical isoelectric point of 6.2. The secondary structure of GBl(2)efl was predicted from the identification of random coils (56.08%), alpha helices (22.22%), extended strands (17.99%), and beta turns (3.7%) through sequence analyses. A homology analysis revealed that GBl(2)efl exhibited a high similarity with other species at the amino acid level, ranging from 52% to 69%. While GBl(2)efl mRNA expression was higher in the dorsal longitudinal flight muscle following a three-day starvation and in the Malpighian tubules following a one-day starvation, no changes in expression were detected in other tissues. Furthermore, tunicamycin-induced endoplasmic reticulum (ER) stress resulted in an approximately 1.8-fold higher expression in the fat body compared with the wild type.

• Journal of Life Science
• /
• v.32 no.9
• /
• pp.721-728
• /
• 2022

This study investigated the diversity of intestinal microbial communities isolated from the intestine of topshell (Turbo cornutus) from the coast of Jeju Island (Beobhwan, Seogwipo city). Pure cultivation using the standard marine agar (MA) medium showed the most significant number of clusters. Aerobic and anaerobic culture allowed isolation of strains of 1.8×10⁵ CFU·g^-1 and 0.4×10 CFU·g^-1 on average, respectively. The microbial population in the topshell intestine was classified into 4 phyla, 12 families, 26 genera, and 67 species. The microbes in the topshell intestine were detected by homology with 93~100% with standard strains. The microbes in the topshell intestine consisted of Proteobacteria 39%, Firmicutes 34%, Actinobacteria 21%, and Bacteroidets 6%. The identified families were Alteromonadaceae (1), Shewanellaceae (4), Vibrionaceae (12), Phyllobacteriaeceae (1), Rhodobacteraceae (8), Bacillaceae (21), Paenibacillaceae (2), Cellulomonadaceae (1), Mycobacteriaceae (6), Nocardiaceae (4), Streptomycetaceae (3) and Flavobacteriaceae (4). Bacillus sp. and Vibrio sp. accounted for the greatest portion of the separated strains. Among the isolated microorganisms, some strains had probiotic functions.

• Korean Journal of Ecology and Environment
• /
• v.55 no.3
• /
• pp.212-220
• /
• 2022

An estuary is a water ecosystem with a high abundance of the species diversity, due to a variety of complex physicochemical factors of the area where freshwater and ocean mixed. The identification of Corbicula species in the estuary environments is difficult because of various morphological characteristics. In this study, we provide taxonomic information on Corbicula species with taxonomic difficulties using morphological and genetic analysis. This study was conducted on clams from the Seomjin River-Gwangyang Bay, one of the major production area of marsh clam in Korea. As a result, we characterized Cytocrome C Oxidase subunit I (COI) sequences of the Corbicula. The 636 bp nucleotide sequences of COI have 98% homology among Corbicula species collected from 2 sites of Seomjin River-Gwangyang Bay. The phylogenetic analysis with 17 species of Corbicula indicated that most of the species collected from Seomjin River-Gwangyang Bay were brackish water clam (Corbicula japonica), and only one Asian clam (Corbicula fluminea). The evolutionary distance between C. japonica and C. fluminea was less than 0.003. Therefore, it was confirmed that C. japonica is phylogenetically closely related to C. fluminea. In 9 species of Cyrenidae, phylogenetic tree was classified into three lineages. These results will be used as an important data for an identification of clam species by providing genetic information for Corbicula species with a morphological diversity.

• Proceedings of the KSAR Conference
• /
• 2001.03a
• /
• pp.52-52
• /
• 2001

Many nucleoside diphosphate (NDP) kinases are ubiquitous enzymes responsible for the exchange of ${\gamma}$-phosphates between tri- and diphosphonucleosides. The catalytic Many nucleoside diphosphate (NDP) kinases are ubiquitous enzymes responsible for the exchange of ${\gamma}$-phosphates between tri- and diphosphonucleosides. The catalytic reaction follows a ping-pong mechanism in which the enzyme is transiently phosphorylated on a histidine residue conserved in all nucleoside diphosphate kinases. Beside their role in nucleotide synthesis, these enzymes present additional functions, possibly independent of catalysis, in processes such as differentiation, cell growth, tumor progression, metastasis and development. To clone murine nm23-M5, several expressed sequence tags (ESTs) of the GenBank data base, selected according to their homology to nm23-H5 cDNA, reconstituted a complete open reading frame (GenBank AF222750). To test whether murine NDPKs (1, 2, 3, 4, 5, and 6) can inhibit Bax-mediated toxicity in yeast, co-transformation was performed respectively. The yeast S.cerevisiae was transformed with a copy expression plasmid containing the histidine selection marker and expressing murine Bax under the control of a galactose-inducible promoter. Several clones were selected and found to be growth inhibited when Bax expression was induced with galactose. A representative clone was transformed again with a copy expression plasmid containing the tryptophane selection marker and expressing either murine Bcl-xL or NDPK under the control of a galactose-inducible promoter. Several subclones of the double-transformants were selected and characterized. The ability of Bcl-xL and NDPKs to suppress Bax-mediated toxicity was determined by growing yeast cells overnight in galactose media and spot-testing on galactose plates starting with an equal number of yeast cells as determined by taking the OD$_{600}$. Ten-fold serial dilutions were used in the spot-test. Plates were grown at 3$0^{\circ}C$ for 2-3 days. All murine NDPKs suppressed Bax dependent apoptosis. Futher study will be peformed whether Bax-toxicity inhibition was caused by NDP kinase activity or additional function.n.

• Korean Journal of Poultry Science
• /
• v.20 no.4
• /
• pp.177-188
• /
• 1993

This study was performed to find out the reasonable sexing methods In the chicken, obtain the basic information for the mechanisms related to chicken sexual differentiation and identify the genes which known to involved in chicken sex differentiation. The chromosome analysis of chicken embryonic fibroblast was a simple method to determine sex of chicken by means of Z and W chromosome identification. The bands of female chicken genomic DNA digested with Xho Ⅰ and Eco RI restriction endonuclease showed to be useful in direct sex determination and these repetitive sequences of Xho Ⅰ and Eco RI families were proposed to be very homologous in their sequences by colony hybridization analysis. Seven of 150 random primers were selected to amplify the W chromosome-specific band by using arbitrary primed PCR and three of them were useful to identify the sex of chicken. To identify the sex differentiation genes in the chicken, PCR for the amplification of ZFY and SRY sequences was performed. ZFY and SRY sequences were amplified successfully in the chicken genome, implying that chicken genome might have the sex-related conserved sequences similar to mammalian ones. The PCR products of ZFY amplification were the same in both sexes, suggesting that these sequences may be located on autosome or Z chromosome. The profile of PCR amplification for SRY sequences showed variation between sexes, but this result was not enough to specify whether the SRY gene in chicken is on the autosome or sex chromosome.

• Journal of Dairy Science and Biotechnology
• /
• v.41 no.4
• /
• pp.191-202
• /
• 2023

In this study, lactic acid bacteria (LAB) was isolated from blueberries. The isolated LAB were rod-shaped and gram-positive, as shown using gram staining. In addition, the identified bacteria showed high homology to Lactiplantibacillus argentoratensis. The culture supernatant was isolated from L. argentoratensis and its antibacterial activity against the pathogenic bacteria Salmonella and Escherichia was analyzed. Culture supernatants of L. argentoratensis significantly inhibited the growth of Salmonella. Enteritidis NCCP 16947, Salmonella Typhimurium NCCP 16960, and Salmonella. Thompson NCCP 11704. Interestingly, the higher the concentration of the culture supernatant, the more significant was the antibacterial activity. Additionally, the culture supernatant of L. argentoratensis showed significant antibacterial activity against Escherichia strains. To determine whether the antibacterial substance is stable to heat and pH, the LAB culture supernatant was heat-treated under 65℃ for 30 min, 75℃ for 15 min, 85℃ for 10 min, and 100℃ for 5 min. Measurement of antibacterial activity against pathogenic strains by adding 5% of heat-treated culture medium showed the same antibacterial activity as before heat treatment. However, in a test where the pH of the culture supernatant was adjusted to 7.0 from 3.73, no antibacterial activity was observed.