• Korean Journal of Ichthyology
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• v.20 no.1
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• pp.1-6
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• 2008

Three types of clonal cell lines were isolated according to their size and phenotype from the adherent cell populations in long-term liquid cultures from the embryonic fibroblast cells of Zebrafish, Danio rerio. All kind of cell lines were well proliferated. The size and number of clonal cell lines derived colonies from stable embryonic cells were significantly increased in the presence of NAC and A2P conditioned medium from the cell lines. The stable cell lines and clonal cell lines were cap-able of well proliferation in vitro. These cell lines have been maintained in continuous culture without change in characteristics. A majority of the clonal cells (80%) was shown a normal chromosomal complement (50 chromosomes, 2N) in according with FACs analysis. Majority of cells were positive to vimentin staining and none of them were positive for nestin and Oct -4 by immunocytochemistry. These results indicate that the clonal cell lines obtained from cultured cells are fibroblasts and may be extremely useful in genetic manipulation for further nuclear transfer and fish cloning.

• Korean Journal of Microbiology
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• v.24 no.3
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• pp.221-227
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• 1986

The strain of Aspergillus nidulans carring a uvsH mutation which had been shown to be absolutely required for UV or 4-NQO induced mutagenic processes was studied on mitotic recombinational behaviour. Although the effect of uvsH locus on spontaneous mitotic crossing over between fpB37 and centromere was not considerable, UV-induced intergenic recombination did not occur in uvsH/uvsH homozygotic diploid. In case of gene conversion at riboflavin locus between a pair of non-complementary alleles, riboA1 and riboA3, the uvsH mutation was not concerned with that process occurred spontaneously or induced by UV irradiation. When the cells were irradiated by UV light, high degrees of aneuploid productions were detected in diploid homozygous for uvsH as compared with wild type, while much difference was not found during normal growth.

• Applied Microscopy
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• v.32 no.4
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• pp.385-398
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• 2002

The purpose of the present study is to investigate whether stromal cells supporting specific microenvironment for hematopoiesis of bone marrow are affected by toxicants and therapeutic drugs such as antibiotics and anticancer drugs and whether laminin-1 is associated with such effects. SD rats were intraperitoneally injected with 75 mg/kg of cyclophosphamide which is widely used to treat infant's solid tumor, leukemia and myeloma and sacrificed after 3 days, 1 week, 3 weeks or 5 weeks of injection. The bone marrow extracted and paraffin-sectioned was analyzed using immunohistochemical staining. A part of tissues was subjected to electron microscopy following reaction with rabbit anti-laminin antibody, biotinylated goat anti-rabbit IgG conjugated with 12 nm gold particles, and staining with uranyl acetate. 1. The bone marrow tissue at day 3 post injection with cyclophosphamide displayed dilated venous sinus, partial necrotic death, and decreased number of hematopoietic cells. Laminin-1 was intensively stained in the reticular and adipose tissues. 2. Up to 5 weeks post injection, laminin-1 was stained at a low level in the stromal tissue of bone marrow and the number of hematopoietic cell was increased. 3. Deposition of the gold particle which represents laminin-1 expression was observed at the highest level in the stromal cells of bone marrow obtained 3 days after injection, and decreased after 1 to 5 weeks. These results suggest that stromal cells which play a role in supporting microenvironment for bone marrow hematopoiesis augment induction of laminin-1 expression and activation upon administration of cyclophosphamide.

• Development and Reproduction
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• v.2 no.1
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• pp.45-52
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• 1998

An enhncer detector line(EDL) having P[1ArB] insertion in X chromosome with expression of reporter gene (lacZ) in the polar cells and border cell of egg chamber was established and used to monitor the differentiation and migration of border cells during the oogenesis of Drosophila. differentiation of border cell from the anterior polar follicle cells was evident in stage-9 egg chamber of EDL149 which was characterized by migration of columnar follicle cells toward posterior of egg chamber surrounding the oocyte. Migration of border cells was observed in the stage-9 and -10 egg chambers. \beta -galactosidase activities were rapidly increased during the first 4 days after eclosion, and it coincided with the timing of border cell differentiation in the ovary during adult life. Homozygote of EDL149 showed some retardation of border cell migration , resulting absence of migration of some border cells in the anterior part of egg chamber or delayed migration of some border cells in the stage-10 egg chamber. These results suggest that the P[1ArB] of EDL149 is inserted at the locus of the structural gene required for the border cell migration. In addition to the expression in egg chambers, lacZ expression was also detected in the meiotic germ cells of testis and antenna, suggesting the possible requirement of the trapped gene function in these organ. this EDL and enhancer trapped gene might be useful for the study of developmentally regulated cell migration.

• Journal of Life Science
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• v.25 no.1
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• pp.84-92
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• 2015

Interferon inducible transmembrane protein (IFITM) family genes have been implicated in various cellular processes such as the homotypic cell adhesion functions of IFNs and cellular anti-proliferative activities. The present study aimed to investigate whether the polymorphisms of the IFITM2 and IFITM5 genes are associated with susceptibility to UC. We identified a total of thirteen polymorphisms (eleven SNPs and two variations) in the IFITM2 gene and twelve polymorphisms (eleven SNPs and one variation) in the IFITM5 gene, by the direct sequencing method. Genotype analysis in the IFITM2 and IFITM5 SNPs was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and Taq-Man probe analysis, and the haplotype frequencies of IFITM2 and IFITM5 SNPs for multiple loci were estimated using the expectation maximization (EM) algorithm. The genotype and allele frequencies of IFITM2 SNPs, as well as IFITM5 SNPs, in UC patients were not significantly different from those of the healthy controls. We also analyzed the combined frequencies of rs77537847 of IFITM1, rs909097 of IFITM2, and rs56069858 of IFITM5 in the UC patients and the healthy controls. Although the distribution of the major combined genotype frequency did not differ significantly between the healthy controls and the UC patients, the GGT combined frequency in the healthy controls was significantly different from that in the UC patients (P=0.002). This result suggests that the combined genotype of the IFITMs polymorphisms may be associated with a susceptibility to UC and could be a useful genetic marker for UC.

• Journal of Korean Medicine Rehabilitation
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• v.15 no.1
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• pp.67-75
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• 2005

목 적 : 면역 세포에서 주로 생성되는 사이토카인인 인터루킨-1 (interleukin-1)은 다양한 신경내분비계와 대사 기능에 영향을 미치며, 비만인의 말초혈액 단핵구에서 증가하는 경향이 있는 것으로 연구보고 되고 있다. 본 연구의 목적은 $IL-1{\alpha}$ 다형성과 사상체질이 비만과 관련이 있는지를 조사하는데 있다. 연구방법 : 본 연구에서는 현저한 체질량지수 (body mass index, BMI) 다양성을 보이는 건강한 한국 여성 182명을 대상으로 $IL-1{\alpha}$ 유전자형을 분석하였다. 결 과 : T 대립 유전자는 BMI 감소와 관련이 있었다. BMI $27-30kg/m^2$ 그룹(group III)에서 T 대립유전자의 빈도는 $25kg/m^2$ 미만의 그룹(group I)에 비하여 유의하게 감소하였다 (odds ratio, OR, 0.141; 0.25% CI, 0.04-0.54). 더욱이 태음인 여성에서 $IL-1{\alpha}$ T 대립유전자의 빈도가 normal group에 비하여 overweight group (BMI $27-29kg/m^2$)에서 명확하게 감소하였다 (OR, 0.139; 95% CI, 0.04-0.54). 결 론 : 본 연구에서 175명의 체질분석결과 $BMI{\geq}25$인 군들의 태음인의 빈도가 높았으며, BMI는 CC genotype를 가진 사람에서보다 CT+TT genotype를 가진 사람에게서 더 낮았다. 태음인에서 동형접합 또는 이형접합 T 대립유전자가 $BMI{\geq}25$인 군에서 유의하게 감소하였다. 이상의 결과는 한국 여성에서 $IL-1{\alpha}$ 유전자 조절부위에 위치한 -889C/T 다형성과 비만 및 사상체질의 관련되는 것을 의미한다.

• Journal of Plant Biotechnology
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• v.43 no.1
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• pp.12-20
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• 2016

Advances in DNA sequencing technologies have contributed to revolutionary understanding of many fundamental biological processes. With unprecedented cost-effective and high-throughput sequencing, a single laboratory can afford to de novo sequence the whole genome for species of interest. In addition, population genetic studies have been remarkably accelerated by numerous molecular markers identified from unbiased genome-wide sequences of population samples. As sequencing technologies have evolved very rapidly, acquiring appropriate individual plants or populations is a major bottleneck in plant research considering the complex nature of plant genome, such as heterozygosity, repetitiveness, and polyploidy. This challenge could be overcome by the old but effective method known as haploid induction. Haploid plants containing half of their sporophytic chromosomes can be rapidly generated mainly by culturing gametophytic cells such as ovules or pollens. Subsequent chromosome doubling in haploid plants can generate stable doubled haploid (DH) with perfect homozygosity. Here, classical methodology to generate and identify haploid plants or DH are summarized. In addition, haploid induction by epigenetic regulation of centromeric histone is explained. Furthermore, the utilization of haploid plant in the genomics era is discussed in the aspect of genome sequencing project and population genetic studies.

• Journal of Life Science
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• v.29 no.5
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• pp.537-544
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• 2019

Genome editing technology employing gene scissors has generated interest in molecular breeding in various fields, and the development of the third-generation gene scissors of the clustered, regularly interspaced short palindromic repeat (CRISPR) system has accelerated the field of molecular breeding through genome editing. In this study, we analyzed the possibility of silkworm molecular breeding using gene scissors by genomic and phenotypic analysis after editing the biogenesis of lysosome-related organelles (BmBLOS) gene of Bakokjam using the CRISPR/Cas9 system. Three types of guide RNAs (gRNA) were synthesized based on the BmBLOS gene sequence of Bakokjam. Complexes of the prepared gRNA and Cas9 protein were formed and introduced into Bombyx mori BM-N cells by electroporation. Analysis of the gene editing efficiency by T7 endonuclease I analysis revealed that the B4N gRNA showed the best efficiency. The silkworm genome was edited by microinjecting the Cas9/B4N gRNA complex into silkworm early embryos and raising the silkworms after hatching. The hatching rate was as low as 18%, but the incidence of mutation was over 40%. In addition, phenotypic changes were observed in about 70% of the G0 generation silkworms. Sequence analysis showed that the BmBLOS gene appeared to be a heterozygote carrying the wild-type and mutation in most individuals, and the genotype of the BmBLOS gene was also different in all individuals. These results suggest that although the possibility of silkworm molecular breeding using the CRISPR/Cas9 system would be very high, continued research on breeding and screening methods will be necessary to improve gene editing efficiency and to obtain homozygotes.