This study was conducted to investigate the effect of feed types on Ochratoxin A (OTA) degradation by Korean native goats. Rumen fluid from canulated goats fed whole roughage or 50% roughage served as a source of micro-organisms. Experiments were undertaken i) to investigate OTA degradation ability in a $2{\times}4$ factorial arrangement with different feed types (100% roughage vs. 50% roughage) and rumen fluid fractions (whole rumen fluid, cells, autoclaved rumen fluid and supernatant) supplemented with OTA ii) to evaluate OTA degradation by the rumen fluid of goats fed two different diets at different time points (0, 3, 6, 9 and 12 h) of feeding iii) to isolate potential rumen microorganisms and iv) to identify elements responsible for OTA degradation. Rumen fluid from goats fed 100% roughage had higher (p<0.05) OTA degradability than 50% roughage diets. OTA degradation based on rumen fluid collection times showed that rumen fluid at 0 h showed significantly higher (p<0.05) degradability. Carboxypeptidase A (CPA) enzyme has been reported to be responsible for OTA degradation. Thus, using real time PCR, primers designed to target the CPA gene from Bacillus licheniformis could be amplified using genomic DNA from rumen fluid of goats and sequenced, thus enabling evaluation of the Bacillus population under different feeding condition and times. Our findings showed that the Bacillus population was significantly higher (p<0.05) before feeding (0 h) in animals which were fed a whole roughage diet, giving indirect evidence of OTA degradation being influenced by Bacillus sps. Thus, it can be concluded that OTA degradability is influenced by feed, feeding time and Bacillus licheniformis population.
Site-specific incorporation of unnatural amino acids into proteins in vivo can be achieved by co-expression of an orthogonal pair of suppressor tRNA and engineered aminoacyl-tRNA synthetase (ARS) that specifically ligates an unnatural amino acid to the suppressor tRNA. As a step to establish this technique, here we generated an Escherichia coli reporter strain DH10B(Tn:lacZam) by integrating amber mutated lacZ gene into the chromosome of E. coli DH10B strain. In vivo expression of E. coli amber suppressor $tRNA^{Gln}$ produced blue colonies in culture plates containing X-Gal as well as dramatically increased $\beta$-galactosidase activity. In addition, expression of an orthogonal pair of Saccharomyces cerevisiae suppressor $tRNA^{Tyr}$ and tyrosyl-tRNA synthetase also produced blue colonies as well as moderate increase of $\beta$-galactosidase activity. These data demonstrate that our reporter strain will provide an efficient method to assess amber suppression in both qualitative and quantitative manners.
In this study, we evaluated the use of RAPD method to discriminate among strains of Fusarium species including F. oxysporum and f. sp. of F. oxysporum. As a result of the amplication, fifteen primers showed total 180 bands ranging from 0.2 to 3 Kb. Among those 180 bands, 126 polymorphic bands were used for bionominal matrix code (0, 1), and UPGMA dendrogram analysis. Fusarium oxysporum isolate 355 showed high similarity with F. oxysporum isolate 358 at 0.9603. Fusarium roseum isolate 87 and F. oxysporum isolate 358, F. o. f. sp. lycopersici isolate 69 and F. o. f. sp. melongena 68 showed low similarity of 0.3809. Fusarium oxysporum isolate 361 and F. o. f. sp. raphani isolate 218 showed similarity of 0.8730, F. oxysoprum isolate 354 and unidentified Fusarium sp. isolate 228 showed similarity matrix of 0.7936, and F. roseum isolate 87 and F. o. f. sp. raphani isolate 57 showed similarity matrix of 0.5873.
Yoon, D. H.;Kim, T. H.;Lee, K. H.;Park, E. W.;Lee, H. K.;Cheong, I. C.;Hong, K. C.
Journal of Animal Science and Technology
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v.45
no.1
/
pp.13-22
/
2003
Growth Hormone (GH) gene is a member of gene family through the evolutionary process from a small common ancestral gene by a series of gene duplications. The role of the GH in growth and performance controls has been extensively studied in human, mice and livestock. Many researchers have considered GH as a strong candidate gene for evaluation of genetic polymorphisms that could be associated with economic traits in cattle. We report here a novel missense mutation within the exon 5 of the bovine Growth Hormone (bGH) gene. We could amplified 522 bp fragments from eight unrelated Hanwoo cattle by PCR, then, subsequently cloned and sequenced. An Msp I RFLP corresponding to a C to T transition was observed at position 2258 nt. From this result, we could predict a missense mutation (Arg to Trp) at codon 166 in a highly conserved region among many mammals. Codominant Mendelian segregation of the two alleles, Msp I (+) and Msp I (-), was observed in two full-sib F2 families (n = 32, African taurine Bos taurus ${\times}$ African zebu Bos indicus) and eight half-sib Hanwoo families. For the availability of genetic marker, we have performed PCR-RFLP with a large number of individual animals from 15 different cattle breeds (European and Asian taurines, and African indicines). Consideration of breed frequencies of Msp I (-) allele in relation to breed type and their geographic origins, shows higher frequencies in humped breeds or Asian cattle breeds than in humpless or European breeds. This result indicates that the missense mutation can be contributed the functional significance such as the signal transduction through the receptor binding, also may be used as a marker for selection of the economic traits in Hanwoo.
The single cell gel electrophoresis (SCGE) assay is a microelectrophoretic technique for assessments of DNA damage at the level of the individual eukaryotic cell. The SCGE assay, due to its simplicity, sensitivity and need of a few cells, has advantages compared to other genomic damage assays such as sister chromatid exchange, chromosomal aberration and micronucleus test. In this study, investigated were the levels of DNA damage and the repair kinetics in the coelomocytes of Eisenia fetida treated with HgCl2 and ionizing radiation by means of the SCGE assay. For detecting DNA damage and repair in coelomocytes, earthworms (E. fetida) were irradiated with six doses of ${\gamma}$-rays (0, 2.5, 5, 10, 20 and 50 Gy) and in vivo exposed to mercuric chloride at 0, 80 and 160 mg $kg^{-1}$ for 48 hours. Then the Olive tail moments were measured during 0~12 hours after irradiation and 0~72 hours after Hg treatment. The results showed that the more the oxidative stress was induced by mercury and radiation, the longer the repair time was required. Also, the results suggest that the SCGE assay may be used as an important tool for comparison of the sensitivity of different species to oxidative stresses.
Sodhi, Simrinder Singh;Jeong, Dong Kee;Sharma, Neelesh;Lee, Jun Heon;Kim, Jeong Hyun;Kim, Sung Hoon;Kim, Sung Woo;Oh, Sung Jong
Korean Journal of Poultry Science
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v.40
no.3
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pp.223-234
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2013
Poultry industry is abounding day by day as it engrosses less cost of investment per bird as compared to large animals. Poultry have the most copious genomic tool box amongst domestic animals for the detection of quantitative trait loci (QTL) and marker assisted selection (MAS). Use of multiple markers and least square techniques for mapping of QTL affecting quality and production traits in poultry is in vogue. Examples of genetic tests that are available to or used in industry programs are documented and classified into causative mutations (direct markers), linked markers in population-wide linkage disequilibrium (LD) with the QTL (LD markers), and linked markers in population wide equilibrium with the QTL (LE markers). Development of genome-wide SNP assays, role of 42 K, 60 K (Illumina) and 600 K (Affymetrix$^{(R)}$ Axim$^{(R)}$) SNP chip with next generation sequencing for identification of single nucleotide polymorphism (SNP) has been documented. Hybridization based, PCR based, DNA chip and sequencing based are the major segments of DNA markers which help in conducting of MAS in poultry. Economic index-marker assisted selection (EI-MAS) provides platform for simultaneous selection for production traits while giving due weightage to their marginal economic values by calculating predicted breeding value, using information on DNA markers which are normally associated with relevant QTL. Understanding of linkage equilibrium, linkage dis-equilibrium, relation between the markers and gene of interest are quite important for success of MAS. This kind of selection is the most useful tool in enhancing disease resistance by identifying candidate genes to improve the immune response. The application of marker assisted selection in selection procedures would help in improvement of economic traits in poultry.
Kim, Soonhee;Hong, Seungbeom;Kemp, Brian M.;Park, Kiwon;Han, Myunsoo
Analytical Science and Technology
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v.21
no.4
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pp.338-343
/
2008
Extraction of DNA from skeletal material is of great importance in the identification of human remains, but is particularly difficult because the high amount of microbial DNA was often co-extracted with human bone DNA. We found that a phenol/chloroform extraction, followed by ultrafiltration, and cleanup by via the $QIAquick^{(R)}$ PCR purification kit yields higher amounts of human genomic DNA compared with extraction by the column affinity $method^{(R)}$ alone. Ultrafiltration extraction of human DNA from ten exhumed bone samples yielded $0.041-1.120ng/{\mu}L$ DNA (mean = $0.498ng/{\mu}L$ DNA), and purification using the column affinity resulted in $0.016-0.064ng/{\mu}L$ DNA (mean = $0.034ng/{\mu}L$ DNA). Although the STR genotyping by the column affinity method was partially successful, all DNA samples by the ultrafiltration method produced full profiles from the multiplex PCR. The efficiency of STR genotyping was in accordance with the amounts of the human DNA extracted.
The aim of this study was to evaluate the genetic diversity and relationships of Ogye populations in Korea. A total of 243 genomic DNA samples from 6 Ogye population (Yeonsan Ogye; YSO, Animal Genetic Resources Research Center Ogye; ARO, Chungbuk Ogye; CBO, Chungnam Ogye; CNO, Gyeongbuk Ogye; GBO, Seoul National University Ogye; SUO) and 3 introduced chicken breeds (Rhode Island Red; RIR, White Leghorn; LG, Cornish; CN) were used. Sizes of 25 microsatellite markers were decided using GeneMapper Software(v 5.0) after analyzing ABI 3130XL. A total of 153 alleles were observed and the range was 2 to 10 per each locus. The mean of expected and observed heterozygosity and PIC (Polymorphism Information Content) value was 0.53, 0.50, 0.46 respectively. The lowest genetic distance (0.073) was observed between YSO and SUO, and the highest distance (0.937) between the RIR and CBO. The results of clustering analysis suggested 3 clusters (${\Delta}K=7.96$). Excluding GBO population, 5 Ogye populations (YSO, ARO, CBO, CNO, SUO) were grouped in same cluster with high genetic uniformity (0.990, 0.979, 0.989, 0.994, 0.985 respectively). But GBO population was grouped in cluster 1 with low genetic uniformity (0.340). The results of this study can be use to basic data for the genetic evaluation and management of Ogye populations in Korea.
Purpose : The urinary tract infection associated with vesicoureteral reflux(VUR) in children may result in serious complications such as renal scarring, hypertension, proteinuria and end stage renal disease. The purpose of this study was to evaluate the factors affecting renal scar such as age, gender, grade of VUR, and ACE gene polymorphism, and body growth in the patients with and those without renal scar associated with VUR Methods : During the period from January 1994 to July 2000, We had 93 children with urinary tract infection associated with VUR who were admitted to the Department of pediatrics of Chonbuk National University Hospital. The patients were divided into two groups according to follow up 99mTc-DMSA renal scan; patients with renal scar group and those with non-scar group. We analyzed and compared the factors associated with renal scarring between the two groups. Results : There were no significant difference in gender, causative organism, ACE gene polymorphism, height and weight at diagnosis between renal scar group and non-scar group. Fifty four patients were in renal scar group and forty seven of them had VUR. The age at diagnosis was significantly higher in renal scar group (2.48${\pm}$2.64yr) than in non renal scar group (1.26${\pm}$1.83yr). Especially, the infants who were less than 1 year of age with VUR developed relatively more renal scar compared with infants older than 1 tear of age. The incidence of renal scarring showed a direct correlation with the severity of VUR. Conclusion : The factors affecting renal scar formation were age at diagnosis, presence and grade of VUR, but the other factors such as gender, causative organism, ACE gene polymorphism were not associated with renal scarring. Therefore, further evaluation about uropathogenic E coli and foflow up study about body growth associated with severity of renal scar would be necessary. (J. Korean Soc Pediatr Nephrol 5 : 43- 50, 2001)
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
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pp.8-9
/
2017
Soybean yield has been low and unstable in Japan and other areas in East Asia, despite long history of cultivation. This is contrasting with consistent increase of yield in North and South America. This presentation tries to describe perspective of breaking stagnation of soybean yield in East Asia, considering the factors of the different yields between regions. Large amount of rainfall with occasional dry-spell in the summer is a nature of monsoon climate and as frequently stated excess water is the factor of low and unstable soybean yield. For example, there exists a great deal of field-to-field variation in yield of 'Tanbaguro' soybean, which is reputed for high market value and thus cultivated intensively and this results in low average yield. According to our field survey, a major portion of yield variation occurs in early growth period. Soybean production on drained paddy fields is also vulnerable to drought stress after flowering. An analysis at the above study site demonstrated a substantial field-to-field variation of canopy transpiration activity in the mid-summer, but the variation of pod-set was not as large as that of early growth. As frequently mentioned by the contest winners of good practice farming, avoidance of excess water problem in the early growth period is of greatest importance. A series of technological development took place in Japan in crop management for stable crop establishment and growth, that includes seed-bed preparation with ridge and/or chisel ploughing, adjustment of seed moisture content, seed treatment with mancozeb+metalaxyl and the water table control system, FOEAS. A unique success is seen in the tidal swamp area in South Sumatra with the Saturated Soil Culture (SSC), which is for managing acidity problem of pyrite soils. In 2016, an average yield of $2.4tha^{-1}$ was recorded for a 450 ha area with SSC (Ghulamahdi 2017, personal communication). This is a sort of raised bed culture and thus the moisture condition is kept markedly stable during growth period. For genetic control, too, many attempts are on-going for better emergence and plant growth after emergence under excess water. There seems to exist two aspects of excess water resistance, one related to phytophthora resistance and the other with better growth under excess water. The improvement for the latter is particularly challenging and genomic approach is expected to be effectively utilized. The crop model simulation would estimate/evaluate the impact of environmental and genetic factors. But comprehensive crop models for soybean are mainly for cultivations on upland fields and crop response to excess water is not fully accounted for. A soybean model for production on drained paddy fields under monsoon climate is demanded to coordinate technological development under changing climate. We recently recognized that the yield potential of recent US cultivars is greater than that of Japanese cultivars and this also may be responsible for different yield trends. Cultivar comparisons proved that higher yields are associated with greater biomass production specifically during early seed filling, in which high and well sustained activity of leaf gas exchange is related. In fact, the leaf stomatal conductance is considered to have been improved during last a couple of decades in the USA through selections for high yield in several crop species. It is suspected that priority to product quality of soybean as food crop, especially large seed size in Japan, did not allow efficient improvement of productivity. We also recently found a substantial variation of yielding performance under an environment of Indonesia among divergent cultivars from tropical and temperate regions through in a part biomass productivity. Gas exchange activity again seems to be involved. Unlike in North America where transpiration adjustment is considered necessary to avoid terminal drought, under the monsoon climate with wet summer plants with higher activity of gas exchange than current level might be advantageous. In order to explore higher or better-adjusted canopy function, the methodological development is demanded for canopy-level evaluation of transpiration activity. The stagnation of soybean yield would be broken through controlling variable water environment and breeding efforts to improve the quality-oriented cultivars for stable and high yield.
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