• Title/Summary/Keyword: M gene

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Overexpression of the SPP2 gene of saccharomyces cerevisiae and production of antibodiesd to Spp2p

  • Park, Kwang-Hark;Lea, Ho-Zoo;L. Woolford;Kim, Kyung-Hoon
    • Journal of Microbiology
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    • v.33 no.3
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    • pp.201-207
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    • 1995
  • We have previously reported that SPP2 gene product of yeast Saccharomyces cerevisiae is involved in the pre-mRNA splicing. To investigate the rol ein the splicing pathway of the Spp2p protein, the SPP2 gene was overexpressed in Escherichia coli and polyclonal antibodies to Spp2p were generated from rabbits. First, a DNA fragment containing the SPP2 GENE without its promoter was subcloned into an E. coli expression vector, pKK233-3. The resulting recombinant plasmid pBQ14 contained an IPTG inducible tac promoter and the SPP2 structural gene. Overexpression of the SPP2 gene was achieved by additionof 0.1 to 1.0 mM IPTG to a logarithmic culture of E. coli JM103(pBQ14) for 90 min at 37.deg.C. Sequence of N-terminal 15 amino acids of the overproduced protein was well matched to the deduced one from the SPP2 reading frame. Then, polyclonal antibodies were generated from rabbits immunized with gel-purified SppSp protein. These antibodies reacted specifically with the Spp2p protein extracted from yeast cells expressing the SPP2 gene to a great extent. The antibodies could also block the activity of yeast splicing extracts.

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Progesterone Inhibits Luteinizins Hormone $\beta$ Subunit (LHP) Gene Expression in the Rat Pituitary in a Svnergic Manner (프로제스테론은 흰쥐 뇌하수체에서 LH$\beta$유전 발현을 에스트로젠과 상승작용으로 억제한다.)

  • 조병남;성재영
    • The Korean Journal of Zoology
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    • v.37 no.3
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    • pp.377-384
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    • 1994
  • The present study examines the inhibitow effect of progesterone (P) on luteinizing hormone $(LH)\beta$ subunit gene expression in anterior pituitary of ovariectomized, estradiol-treated adult rats. A single injection of P (1mg) further decreased the estradiol-Induced decrease in $LH\beta$ mRNA levels in ovariectomTzed rats in a time-dependent manner. p suppressed UIP mRNA levels at lower doses (0.1 and 1mg), but increased $LH\beta$ mRNA levels 81 a high dose (toms). The inhibitor action of P on $Uf\beta$ mRNA was restored when Ru486, a P receptor antagonist, was administered 1h before P treatment. These data clearly indicate that P inhibits gene expression of $LH\beta$ in the rift pituitary in a swersic manner with estrogen.

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Effects of Scutellaria baicalensis Extracts on Tyrosinase Gene Expression in B16 Melanoma Cells (B16 Melanoma Cell에서 티로시나아제 유전자 발현에 황금(Scutellaria baicalensis) 추출물이 미치는 효과)

  • Cho, Nam-Chul;Bai, Suk;Chin, Jong-Eon
    • The Korean Journal of Food And Nutrition
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    • v.23 no.1
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    • pp.118-123
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    • 2010
  • To estimate the regulatory effects of Scutellaria baicalensis extracts on melanin biosynthesis, we evaluated the regulatory effects of the tyrosinase gene on B16 melanoma cells. The results revealed that methanolic extracts of Scutellaria baicalensis resulted in a high increase in the expression of the tyrosinase gene. Specifically, treatment with extracts at concentrations of $10\;{\mu}g/m{\ell}$ and $100\;{\mu}g/m{\ell}$ resulted in increases in tyrosinase gene expression rates of approximately 231% and 256%, respectively, when compared to the control. Moreover, the solvent fraction layers(methylene chloride, ethyl acetate, butyl alcohol, water) improved the expression of the tyrosinase gene, but to a lesser degree than the methanolic extracts. An MTT assay revealed, that the methanolic extract exhibited very low cytotoxicities at $10\;{\mu}g/m{\ell}$ and $100\;{\mu}g/m{\ell}$. Taken together, the results of this study indicated that the methanolic extracts of Scutellaria baicalensis was a very effective positive regulator of tyrosinase gene expression.

Analysis of C43G mutation in the promoter region of the XIST gene in patients with idiopathic primary ovarian insufficiency

  • Yoon, Sang Ho;Choi, Young Min
    • Clinical and Experimental Reproductive Medicine
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    • v.42 no.2
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    • pp.58-61
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    • 2015
  • Objective: The XIST gene is considered to be an attractive candidate gene for skewed X-chromosome inactivation and a possible cause of primary ovarian insufficiency (POI). The purpose of this study was to investigate whether the XIST gene promoter mutation is associated with idiopathic POI in a sample of the Korean population. Methods: Subjects consisted of 102 idiopathic POI patients and 113 healthy controls with normal menstrual cycles. Patients with the following known causes of POI were excluded in advance: cytogenetic abnormalities, prior chemo- or radiotherapy, or prior bilateral oophorectomy. Genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism analysis. Results: The mean age of onset of ovarian insufficiency was $28.7{\pm}8.5years$ and the mean values of serum luteinizing and follicle-stimulating hormones and estradiol in the POI group were $31.4{\pm}18.2mIU/mL$, $74.5{\pm}41.1mIU/mL$, and $30.5{\pm}36.7pg/mL$, respectively. We found no cytosine to guanine (C43G) variation in the XIST gene in both POI patients and controls. Conclusion: The C43G mutation in the promoter region of the XIST gene was not present in the Korean patients with idiopathic POI in our study, in contrast to our expectation, suggesting that the role of XIST in the pathogenesis of POI is not yet clear.

FHIT Gene Expression in Acute Lymphoblastic Leukemia and its Clinical Significance

  • Malak, Camelia A Abdel;Elghanam, Doaa M;Elbossaty, Walaa Fikry
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8197-8201
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    • 2016
  • Background: To investigate the expression of the fragile histidine triad (FHIT) gene in acute lymphoblastic leukemia and its clinical significance. Materials and Methods: The level of expressed FHIT mRNA in peripheral blood from 50 patients with acute lymphoblastic leukemia (ALL) and in 50 peripheral blood samples from healthy volunteers was measured via RT-PCR. Correlation analyses between FHIT gene expression and clinical characteristics (gender, age, white blood count, immunophenotype of acute lymphoblastic leukemia and percentage of blast cells) of the patients were performed. Results: The FHIT gene was expressed at $2.49{\pm}7.37$ of ALL patients against $14.4{\pm}17.9$ in the healthy volunteers. The difference in the expression levels between ALL patients and healthy volunteers was statistically significant. The rate of gene expression did not significantly vary with immunophenotype subtypes. Gene expression was also found to be correlated with increase of total leukocyte and decrease in platelets, but not with age, gender, immunophenotyping or percentage of blast cells. Conclusions: FHIT gene expression is low in acute lymphoblastic leukemia and could be a useful marker to monitor minimal residual disease. This gene is also a candidate target for the immunotherapy of acute lymphoblastic leukemia.

Effect of Taurine on Sperm Characteristics and Gene Expressions in Fresh Boar Semen during In Vitro Storage

  • 장현용;정혜윤;조명현;정희태;박춘곤;김정익;양부근
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.227-227
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    • 2004
  • The purpose of this study was to investigate the effect of taurine on sperm characteristics and gene expressions(bax and Gpx) in fresh boar semen during in vitro storage. The motility of spermatozoa in Modena, Modana plus taurine 25 mM, Modana plus taurine 50 mM, Modana plus taurine 75 mM and Modana plus taurine 100 mM were 63.1%, 65.1%, 65.3%, 82.5% and 80.8%, respectively. (omitted)

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Hormonal Regulation of Leptin, Resistin, and Plasminogen Activator Inhibitor-1 Gene Expression in 3T3-L1 Adipocytes

  • Lee, Hyun-Jung;Kim, Yang-Ha
    • Preventive Nutrition and Food Science
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    • v.9 no.4
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    • pp.336-341
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    • 2004
  • Leptin, resisitn and PAI-1 (plasminogen activator inhibitor-1) are synthesized and secreted by rodent fat cells and recently postulated to be an important link to obesity. This study was conducted to characterize the hormonal regulation of leptin, resistin, and PAI-1 gene expression in the 3T3-L1 adipocytes. The cells were treated with 0.5 $\mu$M insulin, 1 $\mu$M dexamethasone (Dex), or 0.05 $\mu$M triiodothyronine (T3) for 72 hours. The mRNA levels of each peptide were measured by semi-quantitative RT-PCR. The mRNA level of the leptin-producing ob gene was significantly increased by insulin, Dex, and T3 by 3.2-, 3.1- and 2.7-fold, respectively, compared to the control (p < 0.05). The level of resistin mRNA was increased by insulin, Dex, and T3 by 2.7-, 2.5- and 2-fold, respectively, compared to the control (p < 0.05). Likewise, the level of PAI-1 mRNA was significantly increased by insulin, Dex, and T3 compared to the control (p < 0.05). Taken together, our results suggest that insulin, Dex, and T3 may regulate the gene expression of leptin, resistin, and PAI-1 in 3T3-L1 adipocytes.

Molecular Phylogenetic Classification of Dermatophytes Isolated from Dogs and Cats (개와 고양이 유래 피부사상균의 분자생물학적 계통 분석)

  • Kim, Doo;Jeoung, Seok-Young;Ahn, So-Jeo
    • Journal of Veterinary Clinics
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    • v.23 no.4
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    • pp.405-410
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    • 2006
  • Using internal transcribed spacer 1 (ITS1) region ribosomal DNA sequences from 9 strains of Microsporum canis and 5 strains of Microsporum gypseum isolated from dogs and a cat with dermatophytosis, we demonstrated the mutual phylogenetic relationship of these strains. Nucleotide sequence analysis of the ITS 1 gene fragments from the 9 strains of M canis had the 100% nucleotide sequence similarities and the 5 strains of M gypseum also had the 100% nucleotide sequence similarities. The phylogenetic analysis of the nucleotide sequences of the 9 strains of M canis formed a nested cluster with the reference strains of M canis originating from USA, Australia, Japan, and Europe. M canis were genetically distinct from the other reference strains of Microsporum spp, but M distortum, M equinum, and M. ferrugineum were genetically very close to M canis. M gypseum from a cluster in the phylogenetic tree with M canis as an outgroup. The molecular analysis of ITS 1 genes provided the useful information for the identification of these microsporum species and the understanding of their relationship.

Cloning, Expression and Hormonal Regulation of Steroidogenic Acute Regulatory Protein Gene in Buffalo Ovary

  • Malhotra, Nupur;Singh, Dheer;Sharma, M.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.2
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    • pp.184-193
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    • 2007
  • In mammalian ovary, steroidogenic acute regulatory (StAR) protein mediates the true rate-limiting step of transport of cholesterol from outer to inner mitochondrial membrane. Appropriate expression of StAR gene represents an indispensable component of steroidogenesis and its regulation has been found to be species specific. However, limited information is available regarding StAR gene expression during estrous cycle in buffalo ovary. In the present study, expression, localization and hormonal regulation of StAR mRNA were analyzed by semi-quantitative RT-PCR in buffalo ovary and partial cDNA was cloned. Total RNA was isolated from whole follicles of different sizes, granulosa cells from different size follicles and postovulatory structures like corpus luteum and Corpus albicans. Semi-quantitative RT-PCR analyses showed StAR mRNA expression in the postovulatory structure, corpus luteum. No StAR mRNA was detected in total RNA isolated from whole follicles of different size including the preovulatory follicle (>9 mm in diameter). However, granulosa cells isolated from preovulatory follicles showed the moderate expression of StAR mRNA. To assess the hormonal regulation of StAR mRNA, primary culture of buffalo granulosa cells were treated with FSH (100 ng/ml) alone or along with IGF-I (100 ng/ml) for 12 to 18 h. The abundance of StAR mRNA increased in cells treated with FSH alone or FSH with IGF-I. However, effect of FSH with IGF-I on mRNA expression was found highly significant (p<0.01). In conclusion, differential expression of StAR messages was observed during estrous cycle in buffalo ovary. Also, there was a synergistic action of IGF-I on FSH stimulation of StAR gene.

Effects of Sohaphyang-won on the Gene Expression in a Hypoxic Model of Cultured Rat Cortical Cells (배양한 흰쥐 대뇌세포의 저산소증 모델에서 소합향원이 유전자 표현에 미치는 영향)

  • 백진원;이영효;김완식;정승현;신길조;이원철
    • The Journal of Korean Medicine
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    • v.25 no.2
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    • pp.127-137
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    • 2004
  • Objectives : The purpose of this investigation was to evaluate the effects of Sohaphyang-won (SH) on the alteration in gene expression in a hypoxia model using cultured rat cortical cells. Methods : E18 rat cortical cells were grown in neurobasal medium containing B27 supplement. On 12 DIV, SH was added ($20\mu\textrm{g}/ml$) to the culture media for 24 hrs. On 14 DIV, cells were given a hypoxic insult (2% O2/5% CO2, $37^{\circ}C$, 3 hrs), returned to normoxia and cultured for another 24 hrs. Total RNA was prepared from SH-untreated (control) and -treated cultures and alteration in gene expression was analyzed by microarray using rat 5K-TwinChips. Results : Effects on some of the genes whose functions are implicated in neural viability are as follows: 1) For most of the genes altered in expression, the global M values were between -05 to +0.5, Among these, 1517 genes were increased in their expression by more than global M +0.1, while 1480 genes were decreased by more than global M -0.1. 2) The expression of apoptosis-related genes such as Bad (global M =0.35), tumor protein p53 (T53) (global M =0.28) were increased, while v-akt murine thymoma viral oncogene homolog 1 (Akt1) was decreased. 3) The expression of hemoglobin alpha 1 (probably neuroglobin) was increased by about 3.2-fold (global M =1.7). 4) The expression of antioxidation-related catalase gene was increased (global M =0.26). 5) The expression of PKCzeta (prkcz), an upstream kinase of MAPK, was increased (global M =0.29). 6) The expression of retinoic acid receptor alpha (RAR), which may regulate transcription in hypoxic stress, was increased (global M =10.27). Conclusions : In summary, the microarray data suggest that SH doesn't increase the expression of oxygen capture-, anti-oxidation- and 'response to stress' -related genes but decreases some anti-apoptosis genes which would help protect the hypoxic cells from apoptosis.

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