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Regulation of Actin Gene Expression During the Differentiation of Naegleria gruberi

  • Kim, Misook;Lee, Joo-Hun
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.42-48
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    • 2001
  • The regulation of actin gene expression during the differentiation of Naegleria gruberi was examined. Actin mRNA concentration was maximal in amoebae and decreased rapidly after the initiation of differentiation. At 20 min after initiation, the concentration of actin mRNA decreased to 55% of the maximal value. The actin mRNA concentration decreased to the minimum at 80 min (15% of the maximum), and then began to increase slightly at the end of differentiation. This decrease of actin mRNA concentration was regulated by the repression of actin gene transcription based on nuclear run-on transcription experiments. The rates of transcription of actin gene in nuclei prepared at 40 and 80 min after the initiation of differentiation were 50 and 28% of that of nuclei prepared at the beginning of differentiation, respectively. The addition of cycloheximide at the initiation of differentiation inhibited both the rapid decrease in the concentration of actin mRNA and the repression of actin gene transcription. These results suggest that the rapid decrease in the concentration of actin mRNA during the differentiation of N. gruberi is accomplished by the repression of actin gene transcription and this transcriptional regulation requires continuous protein synthesis during the differentiation.

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Effect of Clove Extracts on Tyrosinase Gene Expression (정향(Clove) 추출물이 티로시나아제 유전자의 발현에 미치는 효과)

  • Chin, Jong-Eon;Lee, Jeong-Yong;Kim, Kwan-Chun
    • Journal of environmental and Sanitary engineering
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    • v.22 no.1 s.63
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    • pp.75-83
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    • 2007
  • Clove extract by methanol increased expression of the tyrosinase gene on B16 mouse melanoma cells containing tyrosinase promoter. $10{\mu}g/mL$ and $100{\mu}g/mL$ of the extract showed expression rate of the tyrosinase gene about 138% and 245%, respectively, compared with control. At $500{\mu}g/mL$, expression rate of the extract was impossible to measurement by high cytotoxicity. The solvent fraction of methylene chloride also exhibited highly expression rate as methanol extract. However, the solvent fractions of butyl alcohol and water showed repressive effect on expression of tyrosinase gene at $500{\mu}g/mL$. In MTT assay, cell survival rate of the extract exhibited similar to expression rate of tyrosinase gene. That is, $10{\mu}g/mL$ and $100{\mu}g/mL$ of the extract showed the cell survival rate about 128% and 187%, respectively.

Therapeutic applications of gene editing in chronic liver diseases: an update

  • Shin, Ji Hyun;Lee, Jinho;Jung, Yun Kyung;Kim, Kyeong Sik;Jeong, Jaemin;Choi, Dongho
    • BMB Reports
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    • v.55 no.6
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    • pp.251-258
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    • 2022
  • Innovative genome editing techniques developed in recent decades have revolutionized the biomedical research field. Liver is the most favored target organ for genome editing owing to its ability to regenerate. The regenerative capacity of the liver enables ex vivo gene editing in which the mutated gene in hepatocytes isolated from the animal model of genetic disease is repaired. The edited hepatocytes are injected back into the animal to mitigate the disease. Furthermore, the liver is considered as the easiest target organ for gene editing as it absorbs almost all foreign molecules. The mRNA vaccines, which have been developed to manage the COVID-19 pandemic, have provided a novel gene editing strategy using Cas mRNA. A single injection of gene editing components with Cas mRNA is reported to be efficient in the treatment of patients with genetic liver diseases. In this review, we first discuss previously reported gene editing tools and cases managed using them, as well as liver diseases caused by genetic mutations. Next, we summarize the recent successes of ex vivo and in vivo gene editing approaches in ameliorating liver diseases in animals and humans.

Induction of a Sweetpotato Anion Peroxidase swpa2 Gene Expression by Stress-related Chemicals and Pectobacterium chrysanthemi (스트레스 관련 화합물 처리 및 병원균 감염에 의한 고구마 산성 퍼옥시다제 swpa2 발현 유도)

  • Kim, Yun-Hee;Ryu, Sun-Hwa;Kim, Kee-Yeun;Kwon, Suk-Yoon;Bang, Jae-Wook;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.83-88
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    • 2004
  • Expression of an anionic peroxidase swpa2 gene isolated from cultured cells of sweetpotato (Ipomoea batatas) was investigated under various stress conditions by RT-PCR. The swpa2 gene was not expressed in any tissues of intact sweetpotato plant grown at the normal condition. The expression of this gene was strongly induced in leaf tissue by treatment of $H_2O$$_2$ (440mM). Treatment of NaCl (100mM), ABA (0.1mM) and methyl jasmonate(MeJA, 0.1mM) also induced the expression of swpa2 gene. Interestingly, salicylic acid (SA, 0.1 mM) did not induce the expression of swpa2 gene, indicating that anionic swpa2 POD is differently involved in SA and MeJA signaling pathways. In addition, swpa2 gene was strongly induced in sweetpoato leaf tissues infected with Pectobacterium chrysanthemi, indicating that swpa2 is involved in defense related to the pathogenesis of P. chrysanthemi in sweetpotato plants. These results strongly suggest that swpa2 gene is involved in overcoming oxidative stresses caused by both abiotic and biotic stress.

HisCoM-PAGE: software for hierarchical structural component models for pathway analysis of gene expression data

  • Mok, Lydia;Park, Taesung
    • Genomics & Informatics
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    • v.17 no.4
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    • pp.45.1-45.3
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    • 2019
  • To identify pathways associated with survival phenotypes using gene expression data, we recently proposed the hierarchical structural component model for pathway analysis of gene expression data (HisCoM-PAGE) method. The HisCoM-PAGE software can consider hierarchical structural relationships between genes and pathways and analyze multiple pathways simultaneously. It can be applied to various types of gene expression data, such as microarray data or RNA sequencing data. We expect that the HisCoM-PAGE software will make our method more easily accessible to researchers who want to perform pathway analysis for survival times.

Nutritional and Tissue Specificity of IGF-I and IGFBP-2 Gene Expression in Growing Chickens - A Review -

  • Kita, K.;Nagao, K.;Okumura, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.747-754
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    • 2005
  • Nutritional regulation of gene expression associated with growth and feeding behavior in avian species can become an important technique to improve poultry production according to the supply of nutrients in the diet. Insulin-like growth factor-I (IGF-I) found in chickens has been characterized to be a 70 amino acid polypeptide and plays an important role in growth and metabolism. Although it is been well known that IGF-I is highly associated with embryonic development and post-hatching growth, changes in the distribution of IGF-I gene expression throughout early- to late-embryogenesis have not been studied so far. We revealed that the developmental pattern of IGF-I gene expression during embryogenesis differed among various tissues. No bands of IGF-I mRNA were detected in embryonic liver at 7 days of incubation, and thereafter the amount of hepatic IGF-I mRNA was increased from 14 to 20 days of incubation. In eyes, a peak in IGF-I mRNA levels occurred at mid-embryogenesis, but by contrast, IGF-I mRNA was barely detectable in the heart throughout all incubation periods. In the muscle, no significant difference in IGF-I gene expression was observed during different stages of embryogenesis. After hatching, hepatic IGF-I gene expression as well as plasma IGF-I concentration increases rapidly with age, reaches a peak before sexual maturity, and then declines. The IGF-I gene expression is very sensitive to changes in nutritional conditions. Food-restriction and fasting decreased hepatic IGF-I gene expression and refeeding restored IGF-I gene expression to the level of fed chickens. Dietary protein is also a very strong factor in changing hepatic IGF-I gene expression. Refeeding with dietary protein alone successfully restored hepatic IGF-I gene expression of fasted chickens to the level of fed controls. In most circumstances, IGF-I makes a complex with specific high-affinity IGF-binding proteins (IGFBPs). So far, four different IGFBPs have been identified in avian species and the major IGFBP in chicken plasma has been reported to be IGFBP-2. We studied the relationship between nutritional status and IGFBP-2 gene expression in various tissues of young chickens. In the liver of fed chickens, almost no IGFBP-2 mRNA was detected. However, fasting markedly increased hepatic IGFBP-2 gene expression, and the level was reduced after refeeding. In the gizzard of well-fed young chickens, IGFBP-2 gene expression was detected and fasting significantly elevated gizzard IGFBP-2 mRNA levels to about double that of fed controls. After refeeding, gizzard IGFBP-2 gene expression decreased similar to hepatic IGFBP-2 gene expression. In the brain, IGFBP-2 mRNA was observed in fed chickens and had significantly decreased by fasting. In the kidney, IGFBP-2 gene expression was observed but not influenced by fasting and refeeding. Recently, we have demonstrated in vivo that gizzard and hepatic IGFBP-2 gene expression in fasted chickens was rapidly reduced by intravenous administration of insulin, as indicated that in young chickens the reduction in gizzard and hepatic IGFBP-2 gene expression in vivo stimulated by malnutrition may be, in part, regulated by means of the increase in plasma insulin concentration via an insulin-response element. The influence of dietary protein source (isolated soybean protein vs. casein) and the supplementation of essential amino acids on gizzard IGFBP-2 gene expression was examined. In both soybean protein and casein diet groups, the deficiency of essential amino acids stimulated chickens to increase gizzard IGFBP-2 gene expression. Although amino acid supplementation of a soybean protein diet significantly decreased gizzard IGFBP-2 mRNA levels, a similar reduction was not observed in chickens fed a casein diet supplemented with amino acids. This overview of nutritional regulation of IGF-I and IGFBP-2 gene expression in young chickens would serve for the establishment of the supply of nutrients to diets to improve poultry production.

Effect of Sopyung-tang Extract on Insulin Secretion and Gene Expression in RIN-m5F Cells (소평탕(消平湯)이 RIN-m5F 세포에서 인슐린 분비 및 유전자 발현에 미치는 영향)

  • Youn, Sung-Sik;Cho, Chung-Sik
    • The Journal of Internal Korean Medicine
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    • v.31 no.1
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    • pp.25-39
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    • 2010
  • Background : At high glucose levels in $\beta$-cells, cell viability and insulin secretion are decreased by glucotoxicity. Sopyung-tang(SPT) had an effect on blood glucose level decrease and antioxidant enzyme activities in streptozotocin-induced diabetic rats. Objectives : This study performed a series of experiment to verify the effects of SPT extract on the cell viability, antioxidant enzyme activities, insulin secretion and insulin mRNA expression at hyperglycemic states of RIN-m5F. Methods : After treatment at various concentrations of SPT added to the RIN-m5F cells, cell viability by MTT assay, free radical-scavenging activity, SOD activity and insulin secretion were measured. Additionally, insulin-related gene expression was measured using real-time RT-PCR. Results : Compared to the control group, SPT extract showed considerable effects on RIN-m5F cell viability, DPPH radical-scavenging activity, superoxide dismutase (SOD) activity, insulin secretion and insulin-related gene expression. Conclusions : This study showed that SPT extract has an effect on $\beta$-cell cell viability, insulin secretion and insulin-related gene expression. Thus, SPT extract may be used for treatment of diabetes and its complications. Further mechanism studies of SPT seem to be necessary on the glucotoxicity and oxidative stress.

Induction of Lysozyme Gene Expression During Involution of Mouse Mammary Gland

  • Lee, M.J.;Han, O.;Back, K.;Choi, Y.J.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.4
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    • pp.462-466
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    • 2001
  • To understand molecular mechanisms of mouse mammary gland involution, clones were isolated by differential screening of a cDNA library. Partial sequences of a clone showed 100% identity to cDNA sequences of mouse lysozyme P gene. Northern analysis was performed to examine expression levels of lysozyme mRNA in mammary gland at several physiological states. Expression of lysozyme gene was induced at involution day 5 compared with lactating stage. High levels of lysozyme mRNA were also detected at virgin tissues. Two types of separate genes, P and M lysozyme, have been known in mouse, and we found that both lysozyme P and M genes were expressed in mammary tissues by reverse transcriptase-polymerase chain reaction. The lysozyme enzyme activity determined by lysoplate assay was also higher in involuted mammary tissues compared with lactating tissues, showing a similar trend to its mRNA levels. Lysozyme is an antimicrobial protein and involved in host defense mechanism. The increase in lysozyme gene expression may help to prevent microbial infection during mammary gland involution at which stage the residual milk in the mammary gland provides good nutritional sources for microbial growth.

Characterization of CaCOP1 Gene in Capsicum annuum Treated with Pathogen Infection and Various Abiotic Stresses

  • Guo, Jia;Seong, Eun-Soo;Wang, Myeong-Hyeon
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.227-233
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    • 2007
  • We characterized a full-length cDNA of CaCOP1 from pepper. Phylogenetic analysis based on the deduced amino acid sequence of CaCOP1 cDNA revealed high sequence similarity to the COP1 gene in Oryza sativa (84% identity). CaCOP1 shares high sequence identity with regulatory protein in Arabidopsis (84%), constitutively photomorphogenic 1 protein in Pisum sativum (81%) and COP1 homolog in Lycopersicon esculentum (79%). CaCOP1 gene exists single copy in the chili pepper genome. Expression of CaCOP1 was reduced in response to inoculation of non-host pathogens. The expression of this gene under abiotic and oxidative stresses was investigated, including 200 mM NaCl, 200 mM mannitol, cold ($4^{\circ}C$), 100 ${\mu}M$ abscisic acid (ABA), and 10 mM hydrogen peroxide ($H_2O_2$). CaCOP1 was induced significantly 3 h after low temperature treatment but not by dehydration or high salinity. Moreover, CaCOP1 was not induced by plant hormone ABA. These observations suggest that CaCOP1 gene plays a role in abiotic stress and may be belong to ABA-independent regulation system.

Cloning and Transctiption of Excherichia coli Cell Division Gene, sep (E. coli 세포분열 유전자 sep의 Cloning 및 Transcription에 관한 연구)

  • ;Walker, James R.
    • Korean Journal of Microbiology
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    • v.22 no.4
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    • pp.235-242
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    • 1984
  • Sep gene, which is one of the cell division genes coding for penicillin binding protein 3 was subcloned from ${\lambda}607sep^{+2}$ to plasmid pBR322. which has a strong promotor such as lac UV5(lacP). It was confirmed that the sep gene cloned to pLJ3 was in the proper orientation for expressionfrom lactose promotor. To analyze the expression efficiency of sep gene within the plasmids newly constructed, sep mRNA was assated by using ${\lambda$\mid$\;607sep^{+2}$ DNA as a probe. Sep mRNA level was increased 25 times in the cells carrying sep gene cloned to pBR322 compared to E. coli C600 which has wild type sep gene within the chromosome instead of plasmed. Furthermore, the cells carrying sep gene cloned to pLJ3 derected the synthesis of about 50 times as much sep mRNA as did cells carrying sep gene cloned to pBR 322, representing that the sep gene was successfully cloned to pLJ3.

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