• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.4
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• pp.669-674
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• 1997

This study was attempted to express human lactoferrin gene that has importance as a functional additive in food industry. Lactoferrin has distinctive antibacterial properties. Also, a number of phy-siological roles have been postulated for the lactoferrin in the modulation of immune and inflamatory responses and as a growth factor. Since it did not show feasible growth inhibition by antimicrobial test against HLF, Pichia pastoris was selected the best lactoferrin expression host. HLF expression plasmid pHIL-SI was integrated into the genomic DNA of P. pastoris GSl15. The integration was confirmed not only with 2.4Kb fragment of HLF gene by PCR(polymerase chain reaction) product, but also with same size of specific signal by southern blotting. Among the various pichica transformants, the JY-1 cell showed a positive response for the expression of HLF by the immunoblotting anaysis. The recombinant HLF protein was started to be secreated at 48hr of culture and reached at the highest secreation level at 96hr.

• ALGAE
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• v.36 no.3
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• pp.165-174
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• 2021

Radicilingua Papenfuss and Calonitophyllum Aregood are two small genera of the family Delesseriaceae that consist of only three and one taxonomically accepted species, respectively. The type species of these genera, Radicilingua thysanorhizans from England and Calonitophyllum medium from the Americas, are morphologically very similar, with the only recognized differences being vein size and procarp development. To date, only other two species were recognized inside the genus Radicilingua: R. adriatica and R. reptans. In this study, we analysed specimens of Radicilingua collected in the Adriatic and Ionian Sea (Mediterranean), including a syntype locality of R. adriatica (Trieste, northern Adriatic Sea), alongside material from near the type locality of R. thysanorhizans (Torpoint, Cornwall, UK). The sequences of the rbcL-5P gene fragment here produced represent the first molecular data available for the genus Radicilingua. Phylogenetic reconstruction showed that the specimens from the Adriatic and Ionian Seas were genetically distinct from the Atlantic R. thysanorhizans, even if morphologically overlapping with this species. A detailed morphological description of the Mediterranean specimens, together with an accurate literature search, suggested that they were distinct also from R. adriatica and R. reptans. For these reasons, a new species was here described to encompass the Mediterranean specimens investigated in this study: R. mediterranea Wolf, Sciuto & Sfriso. Moreover, in the rbcL-5P tree, sequences of the genera Radicilingua and Calonitophyllum grouped in a well-supported clade, distinct from the other genera of the subfamily Nitophylloideae, leading us to propose that Calonitophyllum medium should be transferred to Radicilingua.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.4
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• pp.467-476
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• 2019

Objective: This research was conducted to study the genetic diversity in several Indonesian cattle breeds using microsatellite markers to classify the Indonesian cattle breeds. Methods: A total of 229 DNA samples from of 10 cattle breeds were used in this study. The polymerase chain reaction process was conducted using 12 labeled primers. The size of allele was generated using the multiplex DNA fragment analysis. The POPGEN and CERVUS programs were used to obtain the observed number of alleles, effective number of alleles, observed heterozygosity value, expected heterozygosity value, allele frequency, genetic differentiation, the global heterozygote deficit among breeds, and the heterozygote deficit within the breed, gene flow, Hardy-Weinberg equilibrium, and polymorphism information content values. The MEGA program was used to generate a dendrogram that illustrates the relationship among cattle population. Bayesian clustering assignments were analyzed using STRUCTURE program. The GENETIX program was used to perform the correspondence factorial analysis (CFA). The GENALEX program was used to perform the principal coordinates analysis (PCoA) and analysis of molecular variance. The principal component analysis (PCA) was performed using adegenet package of R program. Results: A total of 862 alleles were detected in this study. The INRA23 allele 205 is a specific allele candidate for the Sumba Ongole cattle, while the allele 219 is a specific allele candidate for Ongole Grade. This study revealed a very close genetic relationship between the Ongole Grade and Sumba Ongole cattle and between the Madura and Pasundan cattle. The results from the CFA, PCoA, and PCA analysis in this study provide scientific evidence regarding the genetic relationship between Banteng and Bali cattle. According to the genetic relationship, the Pesisir cattle were classified as Bos indicus cattle. Conclusion: All identified alleles in this study were able to classify the cattle population into three clusters i.e. Bos taurus cluster (Simmental Purebred, Simmental Crossbred, and Holstein Friesian cattle); Bos indicus cluster (Sumba Ongole, Ongole Grade, Madura, Pasundan, and Pesisir cattle); and Bos javanicus cluster (Banteng and Bali cattle).

• Korean Journal of Environmental Agriculture
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• v.41 no.2
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• pp.115-124
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• 2022

BACKGROUND: Methane is a major greenhouse gas attributed to global warming partly contributed by agricultural activities from ruminant fermentation and rice paddy fields. Methanotrophs are microorganisms that utilize methane. Their unique metabolic lifestyle is enabled by enzymes known as methane monooxygenases (MMOs) catalyzing the oxidation of methane to methanol. Rice absorbs, transports, and releases methane directly from soil water to its stems and the micropores and stomata of the plant epidermis. Methylobacterium species associated with rice are dependent on their host for metabolic substrates including methane. METHODS AND RESULTS: Methylobacterium spp. isolated from rice were evaluated for methane oxidation activities and screened for the presence of sMMO mmoC genes. Qualitatively, the soluble methane monooxygenase (sMMO) activities of the selected strains of Methylobacterium spp. were confirmed by the naphthalene oxidation assay. Quantitatively, the sMMO activity ranged from 41.3 to 159.4 nmol min^-1 mg of protein^-1. PCR-based amplification and sequencing confirmed the presence and identity of 314 bp size fragment of the mmoC gene showing over 97% similarity to the CBMB27 mmoC gene indicating that Methylobacterium strains belong to a similar group. CONCLUSION(S): Selected Methylobacterium spp. contained the sMMO mmoC gene and possessed methane oxidation activity. As the putative methane oxidizing strains were isolated from rice and have PGP properties, they could be used to simultaneously reduce paddy field methane emission and promote rice growth.

• Restorative Dentistry and Endodontics
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• v.47 no.2
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• pp.16.1-16.9
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• 2022

Objectives: This study compared the cyclic fatigue resistance of One Curve (C wire) and F6 Skytaper (conventional austenite nickel-titanium [NiTi]), and 2 instruments with thermos-mechanically treated NiTi: Protaper Next X2 (M wire) and Hyflex CM (CM wire). Materials and Methods: Ten new instruments of each group (size: 0.25 mm, 6% taper in the 3 mm tip region) were tested using a rotary bending machine with a 60° curvature angle and a 5 mm curvature radius, at room temperature. The number of cycles until fracture was recorded. The length of the fractured instruments was measured. The fracture surface of each fragment was examined with a scanning electron microscope (SEM). The data were analyzed using one-way analysis of variance and the post hoc Tukey test. The significance level was set at 0.05. Results: At 60°, One Curve, F6 Skytaper and Hyflex CM had significantly longer fatigue lives than Protaper Next X2 (p < 0.05). No statistically significant differences were found in the cyclic fatigue lives of One Curve, F6 Skytaper, and Hyflex CM (p > 0.05). SEM images of the fracture surfaces of the different instruments showed typical features of fatigue failure. Conclusions: Within the conditions of this study, at 60° and with a 5 mm curvature radius, the cyclic fatigue life of One Curve was not significantly different from those of F6 Skytaper and Hyflex CM. The cyclic fatigue lives of these 3 instruments were statistically significantly longer than that of Protaper Next.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.294-294
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• 2022

To date, there have been no reports on the cloning and characterization of a gene encoding SQS from Amaranthus, although there have been some reports on methods of extracting and purifying squalene from Amaranthus seeds. In this study, we monitored the expression pattern of the amaranth SQS gene in seeds at different developmental stages and in different tissues. The transcript expression pattern of the SQS gene was investigated using total RNA isolated from seeds at different stages of development. There were low levels of SQS transcripts at the early stage of seed development, and the levels remained low until the middle developmental stage. The expression of SQS increased rapidly to reach a peak at the mid-late developmental stage, and then declined dramatically. This pattern of expression was consistent with the results of RT-PCR analyses. All RNA samples generated a fragment of the expected size (183-bp). The amaranth SQS was expressed at low levels during the initial to middle stages of seed development, and its expression level increased at the mid-late development stage. Also The tissue-specific expression of amaranth SQS was determined by quantifying its mRNA in total RNA isolated from the leaves, petioles, stems, and roots of seedlings at the four- and six-leaf stages. Using qRT-PCR and RT-PCR analysis, we detected amaranth SQS transcripts in some of the tissues at the six-leaf stage, but in none of the tissues from plants at the four-leaf stage. SQS transcripts accumulated in almost equal amounts in stems and roots, while a lower level accumulated in leaves and petioles during seedling development at the four- to six-leaf stages. This study provides useful information about the molecular characterization of the SQS clone isolated from grain amaranth. A basic understanding of these characteristics will contribute to further studies on the amaranth SQS.

• Fisheries and Aquatic Sciences
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• v.27 no.6
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• pp.346-355
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• 2024

Groupers are economically important species in the fishery and aquaculture industries in Asian countries. Various species of grouper, including hybrids, have been brought to market even without precise species identification. In this study, we analyzed the structure and expression profile of the gene encoding prolactin (PRL) in the red-spotted grouper Epinephelus akaara based on genomic DNA and cDNA templates. The results showed that the PRL gene consists of five exons encoding an open reading frame of 212 amino acids, including a putative signal peptide of 24 amino acids and a mature structural protein of 188 amino acids. It showed amino acid identities of 99% with Epinephelus coioides, 83% with Amphiprion melanopus, 82% with Acanthopagrus schlegelii, 75% with Oreochromis niloticus, 70% with Coregonus autumnalis, and 67% with Oncorhynchus mykiss, indicating its closer similarity to E. coioides and other groupers but marked distinction from non-teleost PRLs. PRL mRNA expression was detected mostly in the brain, including the pituitary gland, with little expression in other tissues. While the 5-exon structure of the PRL gene of red-spotted grouper and the exon sizes were conserved, the sizes of the introns, particularly the first intron, were markedly different among the grouper species. To examine whether these differences can be used to distinguish groupers of similar phenotypes, exon-primed intron-crossing analysis was carried out for various commercially important grouper species. The results showed clear differences in size of the amplified fragment encompassing the first intron of the PRL gene, indicating that this method could be used to develop species-specific markers capable of discriminating between grouper species and their hybrids at the molecular level.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.10-17
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• 2016

Enzyme immunoassay to analyze specific binding activity of antibody to antigen uses horseradish peroxidase (HRP) or alkaline phosphatase (AP). Chemical methods are usually used for coupling of these enzymes to antibody, which is complicated and random cross-linking process. As results, it causes decreases or loss of functional activity of either antibody or enzyme. In addition, most enzyme assays use secondary antibody to detect antigen binding activity of primary antibody. Enzymes coupled to secondary antibody provide a binding signal by substrate-based color development, suggesting secondary antibody is required in enzyme immunoassay. Additional incubation time for binding of secondary antibody should also be necessary. More importantly, non-specific binding activity caused by secondary antibody should also be eliminated. In this study, we cloned AP isolated from Escherichia coli (E. coli) chromosome by PCR and fused to) hAY4 single-chain variable domain fragment (ScFv) specific to death receptor (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand (TRAIL). hAY4 ScFv-AP expressed in E. coli showed 73.8 kDa as a monomer in SDS-PAGE. However, this fusion protein shown in size-exclusion chromatography (SEC) exhibited 147.6 kDa as a dimer confirming that natural dimerization of AP by non-covalent association induced ScFv-AP dimerization. In several immunoassay such as ELISA, Western blot and immunocytochemistry, it showed antigen binding activity by color development of substrates catalyzed by AP directly fused to primary hAY4 ScFv without secondary antibody. In summary, hAY4 ScFv-AP fusion protein was successfully purified as a soluble dimeric form in E. coli and showed antigen binding activity in several immunoassays without addition of secondary antibody which sometimes causes time-consuming, expensive and non-specific false binding.

• Tunnel and Underground Space
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• v.15 no.4 s.57
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• pp.264-274
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• 2005

The optimum blasting pattern to excavate a quarry efficiently and economically can be determined based on the minimum production cost which is generally estimated according to rock fragmentation. Therefore it is a critical problem to predict fragment size distribution of blasted rocks over an entire quarry. By comparing various prediction models, it can be ascertained that the result obtained from Kuz-Ram model relatively coincides with that of field measurements. Kuz-Ram model uses the concept of rock factor to signify conditions of rock mass such as block size, rock jointing, strength and others. For the evaluation of total production cost, it is imperative to estimate 3-D spatial distribution of rock factor for the entire quarry. In this study, a sequential indicator simulation technique is adopted for estimation of spatial distribution of rock factor due to its higher reproducibility of spatial variability and distribution models than Kriging methods. Further, this can reduce the uncertainty of predictor using distribution information of sample data The entire quarry is classified into three types of rock mass and optimum blasting pattern is proposed for each type based on 3-D spatial distribution of rock factor. In addition, plane maps of rock factor distribution for each ground levels is provided to estimate production costs for each process and to make a plan for an optimum blasting pattern.

• Journal of Aquaculture
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• v.9 no.4
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• pp.445-452
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• 1996

Endogeneous activities of ${\beta}-galactosidase$-like enzyme in various tissues from several finfishes and shellfishes were examined by histochemical analysis based on X-gal staining and by fluorimetric measurement using 4-methylumbelliferyl-${\beta}$-D-galactoside (4-MUG). Species used in this study were 3 freshwater fishes, mud loach (Misgurnus mizolepis), common carp (Cyprinus carpio) and tilapia (Oreochromis niloticus) ; 3 marine fishes, olive flounder (Paralichthys olivaceus), stone flounder (Kareius bicoloratus) and marbled sole (Limanda yokohamae) ; and 4 shellfishes, abalone (Haliotis discus hannai), Pacific oyster (Crassoskra gigas), pearl oyster (Pinctada fucata martensii) and ark shell (Anadara broughtonii). The activities of ${\beta}-galactosidase$-like enzyme in all finfishes examined were significantly different among species, with the wide variations between tissues in a species. In general, the tissues such as kidney, intestine and liver were ones which showed the significantly higher values in 4-MUG fluorimetry and deeper staining patterns in X-gal analysis compared to other tissues. On the other hand, serum and muscle revealed the significantly lower activities than others did, regardless of species. Shellfishes were also found to have endogenous activities of ${\beta}-galactosidase$-like enzyme which were significantly varied depending on both species and organs in a species. Hepatopancreas from all shellfishes examined showed the deepest pattern in X-gal staining and also the highest value in 4-MUG analysis, while activities of ${\beta}-galactosidase$-like enzyme in adductor muscles and mantle muscles from all shellfish species in this study except Pacific oyster were negligible : Pacific oyster had the significant activity of this enzyme in muscle tissues. Putative endogenous lacZ fragment was amplified from both finfishes and shellfishes by polymerase chain reaction (PCR). The molecular size of PCR products was about 510 bp, and there was no difference in size among species examined.