• Asian-Australasian Journal of Animal Sciences
• /
• v.31 no.10
• /
• pp.1581-1590
• /
• 2018

Objective: The aim of this study was to clone alternative splicing isoforms of pig myoneurin (MYNN), predict the structure and function of coding protein, and study temporal and spatial expression characteristics of each transcript. Methods: Alternative splice isoforms of MYNN were identified using RNA sequencing (RNA-seq) and cloning techniques. Quantitative real-time polymerase chain reaction (qPCR) was employed to detect expression patterns in 11 tissues of Large White (LW) and Mashen (MS) pigs, and to study developmental expression patterns in cerebellum (CE), stomach (ST), and longissimus dorsi (LD). Results: The results showed that MYNN had two alternatively spliced isoforms, MYNN-1 (GenBank accession number: KY470829) and MYNN-2 (GenBank accession number: KY670835). MYNN-1 coding sequence (CDS) is composed of 1,830 bp encoding 609 AA, whereas MYNN-2 CDS is composed of 1,746 bp encoding 581 AA. MYNN-2 was 84 bp less than MYNN-1 and lacked the sixth exon. MYNN-2 was found to have one $C_2H_2$ type zinc finger protein domain less than MYNN-1. Two variants were ubiquitously expressed in all pig tissues, and there were significant differences in expression of different tissues (p<0.05; p<0.01). The expression of MYNN-1 was significantly higher than that of MYNN-2 in almost tissues (p<0.05; p<0.01), which testified that MYNN-1 is the main variant. The expression of two isoforms decreased gradually with increase of age in ST and CE of MS pig, whereas increased gradually in LW pig. In LD, the expression of two isoforms increased first and then decreased with increase of age in MS pig, and decreased gradually in LW pig. Conclusion: Two transcripts of pig MYNN were successfully cloned and MYNN-1 was main variant. MYNN was highly expressed in ST, CE, and LD, and their expression was regular. We speculated that MYNN plays important roles in digestion/absorption and skeletal muscle growth, whereas the specific mechanisms require further elucidation.

• Molecules and Cells
• /
• v.38 no.11
• /
• pp.950-958
• /
• 2015

BCS1L gene encodes mitochondrial protein and is a member of conserved AAA protein family. This gene is involved in the incorporation of Rieske FeS and Qcr10p into complex III of respiratory chain. In our previous study, AluYRa2-derived alternative transcript in rhesus monkey genome was identified. However, this transcript has not been reported in human genome. In present study, we conducted evolutionary analysis of AluYRa2-exonized transcript with various primate genomic DNAs and cDNAs from humans, rhesus monkeys, and crabeating monkeys. Remarkably, our results show that AluYRa2 element has only been integrated into genomes of Macaca species. This Macaca lineage-specific integration of AluYRa2 element led to exonization event in the first intron region of BCS1L gene by producing a conserved 3' splice site. Intriguingly, in rhesus and crabeating monkeys, more diverse transcript variants by alternative splicing (AS) events, including exon skipping and different 5' splice sites from humans, were identified. Alignment of amino acid sequences revealed that AluYRa2-exonized transcript has short N-terminal peptides. Therefore, AS events play a major role in the generation of various transcripts and proteins during primate evolution. In particular, lineage-specific integration of Alu elements and species-specific Alu-derived exonization events could be important sources of gene diversification in primates.

• Korean Journal of Environmental Biology
• /
• v.37 no.1
• /
• pp.60-67
• /
• 2019

The objective of this study was to verify the stability of nutrient composition by using herbal medicine by-products as an alternative food source and to examine the growth effect on Protaeria brevitarsis seulensis larvae. As a result of comparing the nutritional components of food source, crude protein, crude fat, and crude ash content, except crude fiber content, was high in both non-fermented and fermented medicinal herbal by-products. Especially, crude protein content was highest. Cadmium, lead, mercury, and other heavy metals were not detected and thus stability as alternative food was confirmed. The growth comparison based on the feeding sources showed no significant difference between the fermented oak sawdust fed control group and the herbal medicine by-products fed laboratory group from week 1 to week 3. The weight of a 4 week larva was 0.137 g in the control group and 0.671 g in the laboratory group and so began to reveal differences at a significant level (p<0.05). As a result of comparing weights of Protaetia brevitarsis seulensis larvae according to the level of herbal medicine by-product addition, HMB40 recorded the heaviest weight in week 7. Statistical analysis showed that there was no significant difference in each body weights of HMB40 and HMB80 at week 5 (p<0.05). These results indicate that if the shipping date of an edible insect is a third instar larva, it arrives at the time of shipment at week 5. Thus feeding HMB40 and HMB80 at the 5th week is the most effective.

• Korean Journal of Animal Reproduction
• /
• v.9 no.2
• /
• pp.133-139
• /
• 1985

An immunoassay may be defined as an analytical procedure involving the competitive reaction between a limiting concentration of specific antibody and two populations of antigen, one of which is labelled or immobillized. The advent of immunoassay has revolutionised our knowledge of reproductive physiology and the practice of veterinary and clinical medicine. Radioimmunoassay (RIA) was the first of these methods to be developed, which meausred the analyte with good sensitivity, accuracy and precision (1,2). The essential components of RIA are:-(i) a limited concentration of antibodies, (ii) a reference preparation, and (iii) an antigen labelled with a radioisotope (usually tritium or iodine-125). Most procedures invelove isolating the antibody-bound fraction and measuring the amount of labelled antigen. Good facilities are available for scintilltion counting, data reduction nd statistical analysis. RIA is undergoing refinement through:-(i) the introduction of new techniques to separate the antibody-bound and free fractions which minimize the misclassification of labelled antigen into these compartments, and the amount of non-specfic binding. (3), (ii) the development of non-extration for the measurement of haptens (4), (iii) the determination of a, pp.rent free (i.e. non-protein bound) analytes (5), and (iv) the use of monoclonal antibodies(6). In 1968, Miles and Hales introduced in important new type of immunoassay which they termed immunora-diometric assay (IRMA) based on t도 use of isotopically labelled specific antibodies(7) in a move from limited to excess reagent systems. The concept of two-site IRMAs (with a capture antibody on a solid-phase, and a second labelled antibody to a different antigenic determinant of the analyte) has enabled the development of more sensitive and less-time consuming methods for the measurement of protein hormones ovar wide concentration of analyte (8). The increasing use of isotopic methos for diverse a, pp.ications has exposed several problems. For example, the radioactive half-life and radiolysis of the labelled reagent limits assay sensitivity and imposes a time limit on the usefulness of a kit. In addition, the potential health hazards associated with the use and disposal of radioactive cmpounds and the solvents and photofluors necessary for liquid scientillation counting are incompatable with the development of extra-laboratory tests. To date, the most practical alternative labels to radioisotopes, for the measurement of analytes in a concentration > 1 ng/ml, are erythrocytes, polystyrene particiles, gold sols, dyes and enzymes or cofactors with a visual or colorimetric end-point(9). Increased sensitivity to<1 pg/ml may be obtained with fluorescent and chemiluminescent labels, or enzymes with a fluorometric, chemiluminometric or bioluminometric end-point. The sensitivity of any immunoassay or immunometric assay depends on the affinity of the antibody-antigen reaction, the specific activity of the label, the precision with which the reagents are manipulated and the nonspecific background signal (10). The sensitivity of a limited reagent system for the measurement of haptens or proteins is mainly dependent upon the affinity of the antibodies and the smalleest amount of reagent that may be manipulated. Consequently, it is difficult in practice to improve on the sensitivity obtained with iodine-125 as the label. Conversely, with excess reagent systems for the measurement of proteins it is theoretically possible to increase assay sensitivity at least 1000 fold with alternative luminescent labels. To date, a 10-fold improvement has been achieved, and attempts are being made to reduce the influence of other variables on the specific signal from the immunoreaction.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.4
• /
• pp.791-807
• /
• 2017

The type II secretion system (T2SS), which transports selected periplasmic proteins across the outer membrane, has rarely been studied in nonpathogens or in organisms classified as Betaproteobacteria. Therefore, we studied Cupriavidus metallidurans (Cme), a facultative chemilithoautotroph. Gel analysis of extracellular proteins revealed no remarkable differences between the wild type and the T2SS mutants. However, enzyme assays revealed that native extracellular alkaline phosphatase is a T2SS substrate, because activity was 10-fold greater for the wild type than a T2SS mutant. In Cme engineered to produce three Ralstonia solanacearum (Rso) exoenzymes, at least 95% of their total activities were extracellular, but unexpectedly high percentages of these exoenzymes remained extracellular in T2SS mutants cultured in rich broth. These conditions appear to permit an alternative secretion process, because neither cell lysis nor periplasmic leakage was observed when Cme produced a Pectobacterium carotovorum exoenzyme, and wild-type Cme cultured in minimal medium secreted 98% of Rso polygalacturonase, but 92% of this exoenzyme remained intracellular in T2SS mutants. We concluded that Cme has a functional T2SS despite lacking any abundant native T2SS substrates. The efficient secretion of three foreign exoenzymes by Cme is remarkable, but so too is the indication of an alternative secretion process in rich culture conditions. When not transiting the T2SS, we suggest that Rso exoenzymes are probably selectively packaged into outer membrane vesicles. Phylogenetic analysis of T2SS proteins supports the existence of at least three T2SS subfamilies, and we propose that Cme, as a representative of the Betaproteobacteria, could become a new useful model system for studying T2SS substrate specificity.

• Korean Journal of Agricultural Science
• /
• v.44 no.4
• /
• pp.477-486
• /
• 2017

This review provides an overview of dietary nucleotides as an alternative to in-feed antibiotics for weaning pigs. Dietary nucleotides are composed of DNA or RNA molecules and are normally contained in protein-rich feed ingredient, brewer's yeast, yeast extract, and milk. Weaning pigs are suffering from several stresses, such as environmental challenges (i.e. crowding, transportation, and feeding). Such stressors can damage the intestinal epithelium and cause an invasion by Escherichia coli, secondary inflammatory responses, and post weaning diarrhea. To overcome weaning disorder, people often use antibiotics which reduce symptoms and boost growth performance. However, since antibiotics were banned due to concerns of antibiotic resistant bacteria, researchers are studying alternative materials to antibiotics. Dietary nucleotides are one of the alternative materials for replacing antibiotics and can be used in abnormal conditions, such as weaning diarrhea, low digestibility, and disease condition. Nucleotides have substances that have important roles in cell division and cell growth, affecting growth performance, intestinal condition, and immunological effect at the weaning stage. However, nucleotides' composition is very different between sources and this aspect makes it difficult to utilize nucleotides at the weaning stage. Therefore, this review paper focuses on i) the characteristics and functions of dietary nucleotides and ii) the effect of dietary nucleotides on the growth performance and immune system of pigs.

• Journal of the Korean Society of Food Culture
• /
• v.33 no.1
• /
• pp.48-54
• /
• 2018

This study was performed to evaluate the quality characteristics of functional Dasik prepared with a mixture of freeze-dried mealworm (Tenebrio molitor) powder and dried pollack powder along with assessment of the general and fatty acid compositions of mealworms. General compositions, except for moisture content of freeze-dried mealworm powder, were higher than those of raw mealworms. The ratios of saturated fatty acids and unsaturated fatty acids of freeze-dried mealworm powder and raw mealworms were 1:3.31 and 1:3.19, respectively. Amounts of oleic acid, which was the most abundant among unsaturated fatty acids, of freeze-dried mealworm powder and raw mealworms were 41.12 and 37.84%, respectively. For color values, greater content of freeze-dried mealworm powder in functional Dasik resulted in lower L and b scores, whereas a value increased. In the case of mechanical properties, greater content of freeze-dried mealworm powder resulted in significant reduction of hardness, chewiness, and gumminess. In the case of sensory evaluation, color, taste, and overall quality of DPMD50, which was prepared with a 1:1 ratio of freeze-dried mealworm powder and dried pollack, were the highest. It was concluded that DPMD50 is a nutritious combination of edible insects and fish for protein fortification for growth and the elderly.

• Molecules and Cells
• /
• v.41 no.8
• /
• pp.733-741
• /
• 2018

Mutations in spliceosome components have been implicated in carcinogenesis of various types of cancer. One of the most frequently found is U2AF1 S34F missense mutation. Functional analyses of this mutation have been largely limited to hematological malignancies although the mutation is also frequently seen in other cancer types including lung adenocarcinoma (LUAD). We examined the impact of knockdown (KD) of wild type (wt) U2AF1 and ectopic expression of two splice variant S34F mutant proteins in terms of alternative splicing (AS) pattern and cell cycle progression in A549 lung cancer cells. We demonstrate that induction of distinct AS events and disruption of mitosis at distinct sub-stages result from KD and ectopic expression of the mutant proteins. Importantly, when compared with the splicing pattern seen in LUAD patients with U2AF1 S34F mutation, ectopic expression of S34F mutants but not KD was shown to result in common AS events in several genes involved in cell cycle progression. Our study thus points to an active role of U2AF1 S34F mutant protein in inducing cell cycle dysregulation and mitotic stress. In addition, alternatively spliced genes which we describe here may represent novel potential markers of lung cancer development.

• Fisheries and Aquatic Sciences
• /
• v.22 no.4
• /
• pp.8.1-8.9
• /
• 2019

A 12-week feeding trial was designed to evaluate the effect of total replacement of fish oil (FO) with terrestrial alternative oils on growth, feed utilization, body composition, hematological parameters, and fillet fatty acid profile of mandarin fish juveniles. Four iso-nitrogenous (56% crude protein) and iso-lipidic (13% crude lipid) practical diets were formulated. A control diet contained 6% FO and three other experimental diets were prepared by replacing FO with linseed oil, soybean oil, and lard (designed as FO, LO, SO, and lard, respectively). Each diet was randomly allocated to triplicate groups of 25 fish ($1.8{\pm}0.03g/fish$) in a circular tank. Complete replacement of FO by three tested alternative oils had no remarkable impact on growth performance, feed utilization efficiency, and morphological and hematological parameters of juvenile mandarin fish. However, daily feed intake was found to be significantly higher for fish fed the SO diet compared with those fed the FO and LO diets. Fish fed LO and SO diets exhibited significantly higher levels of the whole body lipid compared to fish fed diet containing FO. Fillet fatty acid composition reflected dietary fatty acid profile. The highest level of ${\alpha}$-linolenic acid, linoleic acid, and oleic acid was observed in fish fillet fed LO, SO, and lard, respectively. Although the eicosapentaenoic acid level of fish fillet fed diet FO was higher than other treatments, no significant difference was found in docosahexaenoic acid content among all dietary groups. The results of the present study clearly demonstrate that the complete replacement of FO in mandarin fish diets is achievable. These findings are useful in dietary formulation to reduce feed costs without compromising mandarin fish growth.

• Animal Bioscience
• /
• v.35 no.2_spc
• /
• pp.332-346
• /
• 2022

Shortage of protein feed resources is the major challenge to the world farm animal industry. Insects are known as an alternative protein source for poultry. A wide range of insects are available for use in poultry diets. Insect larvae thrive in manure, and organic waste, and produce antimicrobial peptides to protect themselves from microbial infections, and additionally these peptides might also be functional in poultry feed. The feed containing antimicrobial peptides can improve the growth performance, nutrient digestibility, intestinal health, and immune function in poultry. Insect meal contains a higher amount of essential amino acids compared to conventional feedstuffs. Black soldier fly, mealworm, housefly, cricket/Grasshopper/Locust (Orthoptera), silkworm, and earthworm are the commonly used insect meals in broiler and laying hen diets. This paper summarizes the nutrient profiles of the insect meals and reviews their efficacy when included in poultry diets. Due to the differences in insect meal products, and breeds of poultry, inconsistent results were noticed among studies. The main challenge for proper utilization, and the promising prospect of insect meal in poultry diet are also addressed in the paper. To fully exploit insect meal as an alternative protein resource, and exert their functional effects, modes of action need to be understood. With the emergence of more accurate and reliable studies, insect meals will undoubtedly play more important role in poultry feed industry.