• Genomics & Informatics
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• v.16 no.1
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• pp.14-20
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• 2018

Despite the importance of mutation rate, some difficulties exist in estimating it. Next-generation sequencing (NGS) data yields large numbers of single-nucleotide polymorphisms, which can make it feasible to estimate substitution rates. The genetic substitution rates of Hanwoo and Holstein cattle were estimated using NGS data. Our main findings was to calculate the gene's substitution rates. Through estimation of genetic substitution rates, we found: diving region of altered substitution density exists. This region may indicate a boundary between protected and unprotected genes. The protected region is mainly associated with the gene ontology terms of regulatory genes. The genes that distinguish Hanwoo from Holstein in terms of substitution rate predominantly have gene ontology terms related to blood and circulatory system. This might imply that Hanwoo and Holstein evolved with dissimilar mutation rates and processes after domestication. The difference in meat quality between Hanwoo and Holstein could originate from differential evolution of the genes related to these blood and circulatory system ontology terms.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.133-133
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• 2017

Lodging is one of the important constraints in rice production. The lodging destroys the canopy structure, and sharply reduces the capacity of photosynthetic rate and dry matter production. In cereal crops, stem lodging can be classified into two types: stem breaking type and stem bending type. To improve stem lodging resistance, it is important to reveal strong culm traits of superior lodging resistant varieties. There are large varietal differences in parameters associated with the bending moment at breaking (M) and flexural rigidity (FR). The indica variety Takanari possesses large M due to its large section modulus (SM) despite of its small bending stress (BS), while Takanari also has large FR due to its large secondary moment of inertia (SMI) and Young's modulus (YM). To identify quantitative trait loci (QTLs) and the corresponding genes associated with the parameters for M ($=SM{\times}BS$) and FR ($=SM{\times}YM$) should enable to develop lodging resistant varieties, efficiently. In order to identify QTLs for cell wall materials such as cellulose, hemicellulose and lignin associated with BS and YM, a set of Chromosome Segment of Substitution Lines (CSSLs) consisted of 37 lines with chromosome segments of Koshihikari in the genetic background of Takanari were used. Takanari had large M and small BS as compared with Koshihikari. The QTLs for BS were estimated on chromosomes 3, 5, 6, 8, 9, 10, 11 and 12. Koshihikari alleles increased BS in these QTLs. Takanari had a large FR due to its large SMI and YM as compared with Koshihikari. The YM was increased by substitution of the Koshihikari chromosomal segments on chromosomes 2, 10 and 11. Other QTLs estimated on chromosomes 7 and 12 that Koshihikari alleles contributed to the decrease of YM. For lignin, only one major QTL for lignin density was detected on chromosome 11. Hollocellulose densities were increased by the substitution of Koshihikari segments on chromosomes 5 and 11. On the other hand, these were decreased on chromosomes 1 and 3 by substitution of Koshihikari segments. QTLs for cellulose density were estimated on chromosomes 1, 3 and 5 by substitution of Koshihikari segments. For hemicellulose, QTL on chromosome 3 showed that hemicellulose density decreased by the substitution of Koshihikari segment. However, hemicellulose densities on chromosomes 5, 8 and 11 showed the opposite effects. The QTLs for hemicellulose, cellulose, and hollocelulose densities identified on chromosome 5 overlapped with that for bending stress, indicating the positive effect of Koshihikari segment on increasing bending stress. These results suggest that some QTLs for the densities of cell wall materials contribute to increasing bending stress and Young's modulus, and could be utilized to improve the lodging resistance for both types of breaking and bending in rice varieties.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.10
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• pp.1404-1410
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• 2008

This study was performed to clarify whether the variation of stress related heat shock protein 70 (HSP70) (GenBank X68213) gene was associated with the nuclear morphological change of in vitro maturation and in vitro capacitation in oocytes of pig ovaries obtained at the slaughterhouse. The nucleic acid substitution of C to G at the 483rd position was found out in HSP70 K1 (290-512) from X68213. The ovaries were categorized into CC, CG, and GG genotypes using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) (BsiHKA I). After the second in vitro maturation of immature fresh oocytes, the relation of nuclear morphological change in oocytes with the genotype of HSP70 K1 gene was such that the MII ratios of the genotype GG and CG (46.93% and 42.20%, respectively) were significantly higher than that of the CC genotype (10.71%) (p<0.05). With respect to in vitro maturation of frozen-thawed oocytes by an open pulled straw (OPS) method, the percentage of oocytes matured to MII stage of the CG genotype showed a higher trend than CC and GG genotypes. After the in vitro maturation of immature fresh oocytes and frozen-thawed oocytes by the OPS method, the relation of the pronuclei change in oocytes matured in vitro with HSP70 genotype was assessed, and the result showed that the enlarged sperm heads (ESH) of matured fresh oocytes and frozen-thawed oocytes were 80.0% and 60.0% in the CC genotype, respectively. The CC genotype group had a significantly higher rate of ESH than the CG and the GG genotype group (p<0.05). The ratios of polyspermic invasion were not different among HSP70 of the three genotypes. It was considered that the rate of in vitro maturation of fertilized oocytes was expected to differ according to genotype of the stress related gene.

• Journal of Life Science
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• v.14 no.6 s.67
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• pp.963-966
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• 2004

The sequence of ITS (partial 16S ribosomal DNA, complete ITS1, 5.8S ribosomal DNA and ITS2, and partial 28S ribosomal DNA) was analysed by PCR and autosequencing in Sarcodon aspratus. The ITS lenght of S. aspratus was 716 base pair. As this sequence compared with other reports on S. aspratus (ace No AF335110), the sequence variation based on nucleotide deletion and substitution was $1.8\%$. This nucleotide variation rate in same species was very higher than in other species. Also, the sequence varitation rates between this S. aspratus and S. imbricatus, and S. squamus were $8\%\;and\;10\%$, respectively. This results suggested that the high sequence variation of S. aspratus might be caused specific host and inhabitat environment which limited gene flow.

• Molecules and Cells
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• v.27 no.3
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• pp.365-381
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• 2009

The chloroplast DNA sequences of Megaleranthis saniculifolia, an endemic and monotypic endangered plant species, were completed in this study (GenBank FJ597983). The genome is 159,924 bp in length. It harbors a pair of IR regions consisting of 26,608 bp each. The lengths of the LSC and SSC regions are 88,326 bp and 18,382 bp, respectively. The structural organizations, gene and intron contents, gene orders, AT contents, codon usages, and transcription units of the Megaleranthis chloroplast genome are similar to those of typical land plant cp DNAs. However, the detailed features of Megaleranthis chloroplast genomes are substantially different from that of Ranunculus, which belongs to the same family, the Ranunculaceae. First, the Megaleranthis cp DNA was 4,797 bp longer than that of Ranunculus due to an expanded IR region into the SSC region and duplicated sequence elements in several spacer regions of the Megaleranthis cp genome. Second, the chloroplast genomes of Megaleranthis and Ranunculus evidence 5.6% sequence divergence in the coding regions, 8.9% sequence divergence in the intron regions, and 18.7% sequence divergence in the intergenic spacer regions, respectively. In both the coding and noncoding regions, average nucleotide substitution rates differed markedly, depending on the genome position. Our data strongly implicate the positional effects of the evolutionary modes of chloroplast genes. The genes evidencing higher levels of base substitutions also have higher incidences of indel mutations and low Ka/Ks ratios. A total of 54 simple sequence repeat loci were identified from the Megaleranthis cp genome. The existence of rich cp SSR loci in the Megaleranthis cp genome provides a rare opportunity to study the population genetic structures of this endangered species. Our phylogenetic trees based on the two independent markers, the nuclear ITS and chloroplast MatK sequences, strongly support the inclusion of the Megaleranthis to the Trollius. Therefore, our molecular trees support Ohwi's original treatment of Megaleranthis saniculifolia to Trollius chosenensis Ohwi.

• Plant Breeding and Biotechnology
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• v.6 no.4
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• pp.426-433
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• 2018

Plant tissue culture is a technique that has invariably been used for various purposes such as obtaining transgenic plants for crop improvement or functional analysis of genes. However, this process can be associated with a variety of genetic and epigenetic instabilities in regenerated plants, termed as somaclonal variation. In this study, we investigated mutation spectrum, chromosomal distributions of nucleotide substitution types of single-nucleotide polymorphisms (SNPs) and insertions/deletions (InDels) by whole genome re-sequencing between Dongjin and Nipponbare along with regenerated plants of Dongjin from different induction periods. Results indicated that molecular spectrum of mutations in regenerated rice against Dongjin genome ranged from $9.14{\times}10^{-5}$ to $1.37{\times}10^{-4}$ during one- to three-month callus inductions, while natural mutation rate between Dongjin and Nipponbare genomes was $6.97{\times}10^{-4}$. Non-random chromosome distribution of SNP and InDel was observed in both regenerants and Dongjin genomes, with the highest densities on chromosome 11. The transition to transversion ratio was 2.25 in common SNPs of regenerants against Dongjin genome with the highest C/T transition frequency, which was similar to that of Dongjin against Nipponbare genome.

• Microbiology and Biotechnology Letters
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• v.31 no.1
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• pp.36-41
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• 2003

In order to increase the production and secretion rate of mouse salivary $\alpha$-amylase from Saccharomyces cerevisiae, various experiments were attempted. A plasmid pCNNinv (AMY) was constructed by the substitution of ADCl promoter and native signal sequence of mouse salivary $\alpha$-amylase cDNA gene with PRBI promoter and yeast invertase leader sequence, which resulted in 25% increase in the production of $\alpha$-amylase in the culture medium. The respiratory deficient transformant carrying pCNNinv (AMY) were obtained by treating yeast cells with ethidium bromide, and the $\alpha$-amylase activities in the culture brothes of the respiratory-deficient transformants were 5-8 times higher than that of parental wild type strain. $\alpha$-Amylase activity was also increased 3 times when the 0.015% (w/v) of 2-mercaptoethanol was added to the culture medium.

• Journal of Animal Science and Technology
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• v.60 no.5
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• pp.14.1-14.12
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• 2018

Background: Sheep production is a major component of the livestock sector in Ethiopia. The country owing to the large population of 30.70 million estimated numbers of sheep in the country and out of which about 72.14% are females, and 27.86% are males with diverse genetic resources. The real value of indigenous breeds was often under-estimated mostly due to their poor appearance and relatively low productivity. Developing countries in most cases opt for exotic breeds to increase animal productivity through crossbreeding or breed substitution without properly investigating the production potential of the indigenous breeds. The main objective of the research was to identify sheep flock composition and structure, farmers' traits of interest and breeding objective of Wollo highland sheep, and their $F_1$ crossbreed progenies. Results: Smallholder farmers' flock synthesized from breeding ewes, breeding rams, pre-weaned ewe lambs, pre-weaned ram lambs, unproductive ewes, castrated and fattened rams, with the percentage coverage of 29.2, 13.3, 15.5, 16.5, 12.4, and 12.5%, respectively. The maximum number of flock size was 289.0 sheep per flock and higher in the third stratum. The off-take rate percentage of the three strata presented as 21.9% in 1st stratum, 12% in the 2nd stratum, and 16.4% in the 3rd stratum and higher off-take rate recorded in the first stratum. Sheep producer's traits of interest ranked by growth rate (first), body size (second) and marketing value was third rank. Communal breeding (random mating), village based controlled breeding, mixed type and private ram controlled breeding practice were comprised of 39.7, 61.7, 52 and 71.3%, respectively. The percentages of ewes per flock composition were presented as 36.5, 27.1 and 25.5%, respectively in the 3rd stratum, 2nd stratum and 3rd stratum in the order of their importance's. Conclusion: Genetic improvement practices at smallholder sheep producers situation was showing promising outcome with indigenous Washera $F_1$ crossbred lambs and which designated for weaning rate, body size, marketing age, age at first lambing, good temperament and large litter size in the order of their rank. The contemporary breeding practice tendency indicated that, reduced flock size to improve flock productivity via crossbreeding practices.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.32-32
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• 2017

To increase rice production, manipulating plant architecture, especially developing new high-yielding cultivars with erect leaves, is crucial in rice breeding programs. Leaf inclination angle determines the light extinction coefficient (k) of the canopy. Erect leaves increase light penetration into the canopy and enable dense plantings with a high leaf area index, thus increasing biomass production and grain yield. Because of erect leaves, the high-yielding indica rice cultivar 'Takanari' has smaller k during ripening than 'Koshihikari', a japonica cultivar with good eating quality. In our previous study, using chromosome segment substitution lines (CSSLs) derived from a cross between 'Takanari' and 'Koshihikari', we detected seven quantitative trait loci (QTLs) for leaf inclination angle on chromosomes 1 (two QTLs), 2, 3, 4, 7, and 12. In this study, we developed a near-isogenic line (NIL-3) carrying a 'Takanari' allele for increased leaf inclination angle on chromosome 3 in the 'Koshihikari' genetic background. We compared k, dry matter production, and grain yield of NIL-3 with those of 'Koshihikari' in the field from 2013 to 2016. NIL-3 had higher inclination angles of the flag, second, and third leaves at full heading and 3 (- 4) weeks after full heading and smaller k of the canopy at the ripening stage. Biomass at full heading and leaf area index at full heading and at harvest did not significantly differ between NIL-3 and 'Koshihikari'. However, biomass at harvest was significantly greater in NIL-3 than in 'Koshihikari' due to a higher net assimilation rate at the ripening stage. The photosynthetic rates of the flag and third leaves did not differ between NIL-3 and Koshihikari at ripening. Grain yield was higher in NIL-3 than 'Koshihikari'. Higher panicle number per square meter in NIL-3 contributed to the higher grain yield of NIL-3. We conclude that the QTL on chromosome 3 increases dry matter and grain production in rice by increasing leaf inclination angle.

• IMMUNE NETWORK
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• v.1 no.2
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• pp.151-161
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• 2001

Background: Inactivation in p53 tumor suppressor gene through a point mutation and deletion is one of the most frequent genetic changes found in human cancer, with 50% of an incidence. This high rate of mutation mostly suggests that the gene plays a central role in the development of cancer and the mutations detected so far were found in exons 5 to 8. Mutation of p53 locus produced accumulation of abnormal p53 protein, and negative regulation of cell proliferation and transcriptional activation as a suppressor of transformation were lost. In addition, inhibition of its normal cellular function of wild-type by mutant is an important step in tumorigenesis. Method: 4 colon cancer cell lines (SNU C1, C2A, C4, C5) were examined for mutation in exons 5 to 8 of the p53 tumor suppressor gene by PCR-SSCP analysis and expression pattern by western blotting and immunoprecipitation. p53-mediated transactivation ability were examined by CAT assay and base substitution of p53 in SNU C2A cell were detected by DNA sequencing. Results: 1) SNU C2A cell and SNU C5 cell were detected mobility shifts each in exon 5 and exon 7 of p53 gene by the PCR-SSCP method, implicating being of p53 mutation. 2) 3 colon cancer cell lines (SNU C1, SNU C2A, SNU C5) expressed wild type and mutant type p53 protein. 3) In northern blot experiment, SNU C2A and SNU C5 cell expressed high level of p53 mRNA. 4) Results of p53-mediated transactivation in colon cancer cell lines by CAT assay represented only SNU C2A cell has transcriptional activity. 5) DNA sequencing in SNU C2A cell showed missense mutation in codon 179 of one allele, histidine to arginine and wild type p53 in the other allele. Conclusion: Colon cancer cell lines showed correlation with mutation in p53 gene and accumulation of abnormal p53 protein. Colon cancer cell SNU C2A retained p53-mediated transactivation as heterozygous p53 with one mutant allele in 179 codon and the other wild-type allele.