• International Journal of Industrial Entomology and Biomaterials
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• v.39 no.1
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• pp.14-21
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• 2019

Genome editing by CRISPR/Cas9, a third-generation gene scissor in molecular breeding at the genome level, is attracting much attention as one of the breeding techniques of the future. In this study, genetic and phenotypic analysis was used to examine the responsiveness of the Bakokjam variety of the silkworm Bombyx mori to molecular breeding using CRISPR/Cas9 in editing the white egg 2 (w-2) gene. The nucleotide sequence of the w-2 gene was analyzed and three different guide RNAs (gRNA) were prepared. The synthesized gRNA was combined with Cas9 protein and then analyzed by T7 endonuclease I after introduction into the Bm-N silkworm cell line. To edit the silkworm gene, W1N and W2P gRNA and Cas9 complexes were microinjected into silkworm embryos. Based on the results of microinjection, the hatching rate was 16-24% and the incidence of mutation was 33-37%. The gene mutation was verified in the heterozygous F1 generation, but no phenotypic change was observed. In F2 homozygotes generated by F1 self-crosses, a mutant phenotype was observed. These results suggest that silkworm molecular breeding using the CRISPR/Cas9 system is possible and will be a very effective way to shorten the time required than the traditional breeding process.

• Horticultural Science & Technology
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• v.28 no.3
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• pp.442-448
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• 2010

The objectives of this study were to analyze mutant lines of Chinese cabbage ($Brassica$ $rapa$ ssp. $pekinensis$) using gene tagging system (plasmid rescue and inverse polymerase chain reaction) and to observe the phenotypic characteristics. Insertional mutants were derived by transferring DNA (T-DNA) of $Agrobacterium$ for functional genomics study in Chinese cabbage. The hypocotyls of Chinese cabbage 'Seoul' were used to obtain transgenic plants with $Agrobacterium$ $tumefaciens$ harboring pRCV2 vector. To tag T-DNA from the Chinese cabbage genomic DNA, plasmid rescue and inverse PCR were applied for multiple copies and single copy insertional mutants. These techniques were successfully conducted to Chinese cabbage plant with high efficiency, and as a result, T-DNA of pRCV2 vector showed distinct various integration patterns in the transgenic plant genome. The polyploidy level analysis showed the change in phenotypic characteristics of 13 mutant lines was not due to variation in somatic chromosome number. Compared with wild type, the $T_1$ progenies showed varied phenotypes, such as decreased stamen numbers, larger or smaller flowers, upright growth habit, hairless leaves, chlorosis symptoms, narrow leaves, and deeply serrated leaves. The polyploidy level analysis showed the change in phenotypic characteristics of 13 mutant lines was not due to variation in somatic chromosome number. To tag T-DNA from the Chinese cabbage genomic DNA, plasmid rescue and inverse PCR were applied for multiple copies and single copy insertional mutants. Mutants that showed distinct phenotypic difference compared to wild type with 1 copy of T-DNA by Southern blot analysis, and with 2n = 20 of chromosome number were selected. These selected mutant lines were sequenced flanking DNA, mapped genomic loci, and the genome information of the lines is being recorded in specially developed database.

• Journal of Microbiology
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• v.40 no.2
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• pp.173-177
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• 2002

In order to identify Yawowia lipolytica genes induced by serum, cDNA representational difference analysis was performed using a PCR-select CDNA subtraction method. One of the genes cloned from the subtraction was a gene (YIMBFl) homologous to Saccharomyces cerevisiae MBF1 encoding the coactivator multiprotein bridging factor 1. Disruption of YIMBFl revealed that the gene was net essential for viability, and the Ylmbf△ strain did not show any distinct phenotypic change on solid serum medium. In liquid medium, however. a difference was found in the ability to maintain hyphae induced by serum. This result suggests that the YIMbf1 protein may mediate transcriptional activation of certain genes involved in the hypha fonmation of Y. lipolytica.

• Korean Journal of Pharmacognosy
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• v.35 no.1 s.136
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• pp.62-79
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• 2004

Cell-scattering is a phenotypic change easily observed in most epithelial cells treated with Hepatocyte Growth Factor /scatter Factor (HGF/SF) or phorbol esters (PKC-activators). Recent studies have shown the possibilities to use as therapeutic materials of HGF/SF or non tumor promoting phorbol esters for liver disease, cancer and AIDS. In this study, we tested a cell-scattering activity of 534 methanol extracts from plants inhabiting in Korean peninsula using the phenotype-based assay system. Nine Active extracts were detected : Daphne genkwa, Daphne kiusiana, and Aleurites fordii showed high activity (+++), Euphorbia sieboldiana and Rhodotypos scandens showed medium activity (++), Sambucus sieboldiana var. pendula, Catalpa bignonioides, Sambucus sieboldiana and Lycoris squamigera showed low activity (+). Furthermore, the effects of these active materials in the culture cells were investigated with biochemical studies.

• Molecules and Cells
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• v.46 no.1
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• pp.48-56
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• 2023

Genomic information stored in the DNA is transcribed to the mRNA and translated to proteins. The 3' untranslated regions (3'UTRs) of the mRNA serve pivotal roles in post-transcriptional gene expression, regulating mRNA stability, translation, and localization. Similar to DNA mutations producing aberrant proteins, RNA alterations expand the transcriptome landscape and change the cellular proteome. Recent global analyses reveal that many genes express various forms of altered RNAs, including 3'UTR length variants. Alternative polyadenylation and alternative splicing are involved in diversifying 3'UTRs, which could act as a hidden layer of eukaryotic gene expression control. In this review, we summarize the functions and regulations of 3'UTRs and elaborate on the generation and functional consequences of 3'UTR diversity. Given that dynamic 3'UTR length control contributes to phenotypic complexity, dysregulated 3'UTR diversity might be relevant to disease development, including cancers. Thus, 3'UTR diversity in cancer could open exciting new research areas and provide avenues for novel cancer theragnostics.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.5
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• pp.610-615
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• 2011

We examined the correlations between lactation curve shape, including persistency and changes in body condition score (BCS) during early-stage (0 to 30 days in milk (DIM)), nadir-stage (31 to 90 DIM), and late-stage (91 to 240 DIM) lactation in 191 first-lactation cows. Data used were first-parity BCS records, scored twice every month after calving, and daily milk yields. Individual lactation curves were depicted by the Wilmink function. Lactation persistency was defined as the difference in estimated milk yields between 240 DIM and 60 DIM. Changes in BCSs in the early and late stages were defined as linear regression coefficients. There were no significant correlations between traits for lactation curve shape and change in BCS in early-stage lactation. Peak yield and total milk yield were negatively correlated with BCSs in nadir- and late-stage lactation and with BCS change in late-stage lactation, suggesting that cows with high lactation yields had low body reserves and health status in mid- to late lactation and had delayed recovery of body reserves. Lactation persistency was positively correlated with change in BCS in late-stage lactation, suggesting that cows with high lactation persistency tended to be healthy and to recover their body reserves well in late lactation.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.9
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• pp.1259-1265
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• 2015

The present study was performed to study the genetic relationship of productive life with production and type traits of Korean Holsteins at first three lactations. The data for the analysis from 56,054, 28,997, and 11,816 animals of first, second and third parity cows which were born from 2006 to 2011 were collected by Dairy Cattle Improvement Center, National Agricultural Co-operative Federation. Milk, protein and fat yields adjusted for 305 days and average somatic cell score considered as production traits and analyzed type traits were stature, strength, body depth, dairy form, rump angle, rump width, rear leg side view, foot angle, front attachment placement, rear attachment height, rear attachment width, udder cleft, udder depth, front teat placement and front teat length. A multi trait genetic analysis was performed using Wombat program with restricted maximum likelihood animal model composed of fixed effect of birth year, farm and the random effect of animal and random residual effect according to the traits. Heritability estimates of productive life were between 0.06 and 0.13. Genetic and phenotypic correlations between production and productive life traits ranged from 0.35 to 0.04 for milk, 0.16 to 0.05 for protein and 0.18 to 0.02 f 15-0034 (2nd) 150520 or fat. Somatic cells score showed a negative genetic and phenotypic correlation with productive life and also udder type traits, indicating that the selection for higher udder traits will likely to improve resistance to mastitis and persistence in the herd. Among all dairy form type traits, udder characters such as udder cleft showed a significant relationship with productive life. However, a specific change of heritabilities or correlations were not observed with the change of parity. Moreover, further studies are needed to further confirm the significance of the above traits and the effect of parity on above relationships in order to minimize both voluntary and involuntary culling rates while improving herd health and maintaining high yielding dairy cows.

• Journal of Plant Biotechnology
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• v.7 no.2
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• pp.129-134
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• 2005

Callus induction from a leaf explant has been achieved in Lilium Oriental hybrid 'Casa Blanca'. The highest frequency of callus induction was obtained on MS medium supplemented with 0.5 mg/L BA and 2.0 mg/L NAA after 2 months of culture. The cultures maintained continuously without change in color and type of callus when they cultured in the dark. Plantlet regeneration with a high frequency was achieved from induced calli on the same medium. A number of shoots are formed from one cluster of callus, and bulblets developed into intact plantlets after transfer to hormone-free MS medium. No phenotypic variations were observed among regenerants. Enhancement in plantlet regeneration via callus formation would be expected to facilitate the efficiency of transformation of this Oriental hybrid 'Casa Blanca'.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.149-154
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• 2017

Transfer of Micropolyspora internatus into the genus Saccharomonospora as "Saccharomonospora internatus comb. nov." was proposed by Kurup and Greiner-Mai, but the nomenclatural change has not been validly published. Although the inclusion of M. internatus in the genus Saccharomonospora has not being established, the synonymy of "Saccharomonospora internatus" and S. viridis was proposed by Greiner-Mai. A number of recent publications regarded M. internatus as a synonym of S. viridis, but the name M. internatus is still used in some cases instead of S. viridis. This is because of the complicated history of M. internatus and S. viridis, but it is different from the generally accepted view of prokaryotic taxonomy. To clearly verify the synonymy of M. internatus and S. viridis, a literature review and experimental verification were conducted in this study. Based on the genomic and phenotypic characteristics obtained in this study, the synonymy of the two species was obvious. The emended description of S. viridis is given.

• The Korean Journal of Malacology
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• v.27 no.4
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• pp.345-351
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• 2011

Comparison of different morphological characteristics of bivalves is very useful for distinguishing species. Therefore, this study used a morphometric approach to document patterns of phenotypic change through the specimens of family Pinnidae inhabiting the bottom sediments at the coasts of An Thoi Archipelago (south-eastern Gulf of Thailand), in that way excluding intraspecific variations among different populations along a wide geographical range. It was revealed that individuals determined as Pinna trigonium separated from specimens of Pinna nigra and Atrina vexillum at high level, but P. nigra and A. vexillum were the single species A. vexillum, as it is assumed now. Also, it was found that both groups of P. trigonium individuals and A. vexillum specimens divided into tree subgroups unified the specimens with the similar morphometric parameters: small, medium and large within the populations, though there were no evident divisions into subgroups in the size distributions of the populations. In addition to the interspecific morphological variations, the species also showed significant intraspecific morphological variations even in the same population. Most probable reason for the appearance of the intraspecific morphological variations and division of all specimens into size-specific subgroups in the population is the fact that the growth of Pinnidae bivalves is not isometric.