• Journal of the Korean Society of Food Science and Nutrition
• /
• v.19 no.4
• /
• pp.335-341
• /
• 1990

Nuclei proteins were purified from chick liver to homogeneity by means of acid extraction CM Sephadex c 25 column chromatography and Bio Rex 70 column chromatography, The molecular weight of liver Nuclei proteins 1 and 2 as estimated by electrophoresis on SDS-polycrylamide gel are 29000 and 27,000 respectively. These molecular weights are identical with those of Nuclei Proteins 1 and 2 isolated from chick erythrocyte. The liver and erythrocyte Nuclei Proteins also co-migrated in acetic acid-urea gel electrophoresis. Furthermore the anti-sera raised against liver Nuclei Proteins 1 and 2 cross-reacted with erythrocyte Nuclei Proteins 1 and 2 respectively, However the amino acid compositions of liver Nuclei Prooteins 1 and 2 were found to be different from those of corresponding erythrocyte Nuclei proteins ; the contents of serine and proline in liver Nuclei proteins were higherocyte Nuclei proteins ; the contents of serine and proline in liver Nuclei protesins were higher than those in erythrocyte Nuclei proteins while the content of lycsine in liver Nuclei proteins was lower than the erythrocyte Nuclei proteins, These results suggest that in spite of similarities in many respects the liver and erythrocyte Nuclei proteins in chicks and different proteins.

• Microbiology and Biotechnology Letters
• /
• v.18 no.6
• /
• pp.560-565
• /
• 1990

When protoplasts from auxotrophie mutant of Trkhoderma koningii CUT121(Lys-, Met-) were mixed with isolated nuclei of wild type T. koningii ATCC 261 13 and treated with PEG solution, protrophic colonies were produced with frequency of more than 30 percent. One of segregants from prototrophic colonies showed increased xylanase activity and other polysaccharide-hydrolyzing activities comparable to those of wild type strain. Through measurement of DNA contents, induced segregation, and analysis of isozyme patterns, it was revealed that the prototrophic colonies were transformants resulted from exchange of genetic materials between the two kinds of nuclei used. These results suggest that nuclei transfer technique is more efficient than conventional protoplast fusion technique for strain improvement of Trichoderma species.

• The KIPS Transactions:PartB
• /
• v.16B no.2
• /
• pp.131-140
• /
• 2009

This paper presents a new algorithm to the segmentation of the FISH images. First, for segmentation of the cell nuclei from background, a threshold is estimated by using the gaussian mixture model and maximizing the likelihood function of gray value of cell images. After nuclei segmentation, overlapped nuclei and isolated nuclei need to be classified for exact nuclei analysis. For nuclei classification, this paper extracted the morphological features of the nuclei such as compactness, smoothness and moments from training data. Three probability density functions are generated from these features and they are applied to the proposed Bayesian networks as evidences. After nuclei classification, segmenting of overlapped nuclei into isolated nuclei is necessary. This paper first performs intensity gradient transform and watershed algorithm to segment overlapped nuclei. Then proposed stepwise merging strategy is applied to merge several fragments in major nucleus. The experimental results using FISH images show that our system can indeed improve segmentation performance compared to previous researches, since we performed nuclei classification before separating overlapped nuclei.

• Journal of Microbiology and Biotechnology
• /
• v.4 no.3
• /
• pp.183-190
• /
• 1994

Cell-free extracts prepared from cultured insect cells, Spodoptera. frugiperda, were analyzed for activation of early gene transcription of an insect baculovirus, Autographa californica nuclear polyhedrosis virus (AcNPV). The template DNA used for in vitro transcription assays contained promoter sites for the baculovirus genes that have been classified as immediate early ($\alpha$) or early genes. These genes are located in the HindIII-K/Q region of the AcNPV genome. Nuclei isolated from the AcNPV-infected Spodoptera frugiperda cells were also used for in vitro transcription analysis by RNase-mapping the labeled RNA synthesized from in vitro run-on reaction in the isolated nuclei. The genes studied by this technique were p26 and pl0 genes which were classified as delayed early and late gene, respectively. We found that transcription of the genes from the HindIII-K region was accurately initiated and unique in the whole cell extract obtained from uninfected cells, although abundance of the in vitro transcripts was reverse to that of in vivo RNA. With isolated nuclei transcription of the p26 gene was inhibited by $\alpha$-amanitin suggesting that the p26 gene was transcribed by host RNA polymerase II. However, transcription of the pl0 gene in isolated nuclei was not inhibited by $\alpha$-amanitin, but rather stimulated by the inhibitor. We also found that the synthesis of $\alpha$-amanitin-resistant RNA polymerase was begun before 6 hr p.i., the time point at which the onset of viral DNA replication as well as the appearance of a-amanitin-resistant viral transcripts were detected. These studies give us strong evidence to support the previous data that early genes of AcNPV were transcribed by host RNA polymerease III, while transcription of late genes was mediated at least by a novel $\alpha$-amanitin-resistant RNA polymerase.

• Archives of Pharmacal Research
• /
• v.9 no.3
• /
• pp.157-161
• /
• 1986

Isolated calf thymus nuclei were in vitro methylated with S- adenosy-L-methyl-$^{14}C$ methionine, and the proteins were fractionated according to their solubilities. Histone fraction ($H_{2}SO_{4}$-soluble fraction) contained approximately 60% total radioactivity incorporated, while "residual protein" which was ($H_{2}SO_{4}$-insoluble contained the remaining radio-activity. The "residual protein" was further fractionated into various acidic proteins, which contained very littel of the radioactivity. However, the protein fraction eluted from DEAE-cellulose with 0.5 N NaOH contained the largest amount of radioactivity. This protein was found to be basic in nature by amino analysis.

• The Korean Journal of Mycology
• /
• v.16 no.4
• /
• pp.210-213
• /
• 1988

Several reversion colonies were obtained after induced transfer of the isolated nuclei from P. sapidus into protoplast of P. ostreatus$({Arg}^-)$. These colonies showed three distinct cultural characteristics, type 1 produced spontaneous segregants of both parental types, type 2 showed segregants of non parental types, and type 3 gave rise to homogeneous colonies. Isozyme patterns of esterase, malate dehydrogenase and superoxide dismutase showed substantial differences between parents and nuclei transferred strains. This observation supported that the isolated nuclei of P. sapidus were transferred into protoplast of P. ostreatus and expressed in recipient cell.

• Journal of Life Science
• /
• v.8 no.4
• /
• pp.400-409
• /
• 1998

Formation of adduct was studied in benzo(a)pyrene(BP)- and doxorubicin(Dx)-treated human NC-37 cells and isolated nuclei. Major adducts formed were determined by fluorescence absorption spectrophotometery and DNA-lin-ked protein assay. When isolated nuclei were exposed to carcinogens BP and DMBA, and anticancer drugs m-AMSA, ellipticine and Dx, varying degrees of adduct formation occured between DNA-protein complex and these drugs. When the mixture was centrifuged 1.7 M sucrose solution, binding BP and DMBA appeared to be similar between the sediment and the supernatant. When the sediment was centrifuged again with 0.35% polymin-P, the amount of BP bound was 2-fold greater in the protein(1077$\pm$55cpm) than in DNA fraction (470$\pm$20cpm), whereas that of DMBA was 1.6-fold greater in the DNA than in protein fraction. In the case of m-AMSA, ellipticine and Dx, the amount of binding was slightly greater in supernatant than in sediment in centrifugation with 1.7 M sucrose, and more than 3 times greater in the DNA- than in protein- fraction in centrifugation with 0.35% polymin P. DNA fractions which associated with a subset of nonhistone chromosomal protein were isolated from NC-37 cells exposed to $^{3}$H-BP and $^{14}$C-Dx. They were separated into two distince components DNA-S and DNA-P by centrifugation with 2M Nacl chromatin extraction. The results indicated that the amount of $^{3}$H-BP bound was 6.0-fold greater in DNA-P as compared with DNA-S, while that of $^{14}$C-Dx binding appreaed to be 6.2-fold greater in DNA-S than in DNA-P fraction. When $^{3}$H-BP binding wasdetermined in the presence of cold Dx, the amount of binding was reduced only in the DNA-P fraction, indicating that the interaction between DNA and protein is decreased. Gene expression by these drugs, BP treated cells were increased to compare with nomal cells but reduced by treatment with BP-Dx. These results suggest that the protein moiety which tightly bound to DNA-P fraction may play an important role in the regulation of gene expression.

• Korean Journal of Animal Reproduction
• /
• v.16 no.4
• /
• pp.341-346
• /
• 1993

The development of isolated blastomeres from mammalian preimplantation embryos has been basically studied for the multiplication of embryos from superior animals. Therefore, this study was investigated the effect of co-culture with mouse fetal fibroblast cells(MFFC) on in vitro development of blastomeres from mouse preimplantation embryos. Mature female ICR mice were treated with hormone to induce superovulation and embryos were collected at each 2, 4, and 8-cell stage. Then, after removing zona pellucida with protease, blastomeres were isolated by micropipetting, or reconstituted with different stage blastomere, and incubated for 72 hrs either in T6 or TCM199 or on the monolayer of MFFC, which was prepared with fibroblast cells from 14∼14 day mouse fetus. After incubation, we examined their development rates every day and the nuclei numbers of each blastocyst by Hoechst-33342 staining. In the development rates of blastomeres, there were no significant differences between media but the higher rateswere found in the monolayer of MFFC, regardless of reconsititution. In addition, blastomeres cultured with MFFC had slightly greater number of nuclei than those cultured in single media. Generally, the higher development rates of blastomeres were found from earlier stage embryos than the later ones, regardless of culture conditions. Reconsitituted blastomeres had more nuclei but did not show the higher development rates, compared to the single blastomeres. Taken together, our results suggest that co-culture with MFFC have a beneficial effect on the in vitro development of blastomeres from mouse embryos.

• Journal of Microbiology and Biotechnology
• /
• v.7 no.3
• /
• pp.161-166
• /
• 1997

Nuclei were isolated from the protoplasts of Trichoderma harzianum T95 and treated with colchicine, a polyploid inducer. The nuclei were transferred into the protoplast of multi-auxotrophic Gliocladium virens G88 which cannot grow in minimal medium. The protoplast of G. virens G88 carrying the transferred nuclei were regenerated in a regeneration minimal medium containing $17{\mu}g/ml$ of chloroneb as a haploid inducer. Six intergeneric hybrids between G. virens and T. harzianum were isolated from the regeneration minimal medium. The hybrids could be classified into three types according to morphology, those with an isozyme pattern, those with an protein band and those with an randomly amplified polymorphic DNA(RAPD) pattern produced by random primers and repetitive sequences. The first group was identified to be a haploid recombinant, the second group a heterokaryon, and the third appeared to be petite.

• The Korean Journal of Mycology
• /
• v.15 no.4
• /
• pp.250-253
• /
• 1987

The transfer of the isolated nuclei from P. florida into protoplasts of P. ostreatus was induced with polyethylene glycol and $CaCl_2$. Three types of transfer products of nuclei were obtained when transferred to MMM. Type 1 colonies were more vigrously growing mycelium and stable on MCM. One of the type 1 colonies, appeared segregation on MCM plus benomyl. The mycelium did not form clamp connection. These results suggest that type 1 colonies were nuclear hybrids or allodiploids. Type 2 was main products of nuclear transfer. The mycelium formed clamp connection and fertile on sawdust media. Type 3 was very slow growing or non-viable colonies after debris of nuclei or chromosomes transfer into recipient protoplasts. Isozyme pattern of esterase in type 1 produced a new band. Type 2 and type 3 could be characterized by parental bands.