• Title/Summary/Keyword: H2A.Z gene

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Human Estrogen Receptor Ligand Binding Domain (hER LBD)과 Co-activator로 구성된 효모 Two-Hybrid System을 이용한 내분비계장애물질 검출계의 구축 (Construction of the Detection System of Endocrine Disrupters using Yeast Two-Hybrid System with Human Estrogen Receptor ligand Binding Domain and Co-activators)

  • 이행석;조은민;류재천
    • 한국환경성돌연변이발암원학회지
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    • 제22권3호
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    • pp.175-182
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    • 2002
  • Endocrine disruptors (EDs) are the chemicals that affect endocrine systems through activation or inhibition of steroid hormone response. It is necessary to have a good system to evaluate rapidly and accurately endocrine-disrupting activities of suspected chemicals and their degradation products. The key targets of EDs are nuclear hormone receptors, which bind to steroid hormones and regulate their gene transcription. We constructed a co-expression system of Gal4p DNA binding domain (DBD)- ligand binding domain of human estrogen receptor $\alpha$ or $\beta$, and Gal4p transactivation domain (TAD)-co-activator AIB-1, SRC-1 or TIF-2 in Saccharomyces cerevisiae with a chromosome-integrated lacZ reporter gene under the control of CYC1 promoter and Gal4p binding site (GAL4 upstream activating sequence, GAL4$_{UAS}$). Expression of this reporter gene was dependent on the presence of estrogen or EDs in the culture medium. We found that the two-hybrid system with combination of the hER$\beta$ LBD and co-activator SRC-1 was most effective in the xenoestrogen-dependent induction of reporter activity. The extent of transcriptional activation by those chemicals correlated with their estrogenic activities measured by other assay systems, indicating that this assay system is efficient and reliable for measuring estrogenic activity. The data in this research demonstrated that the yeast detection system using steroid hormone receptor and co-activator is a useful tool for identifying chemicals that interact with steroid receptors.s.

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Alcaligenes faecalis NS13에 의한 호기성 종속영양 질산화 및 탈질화 (Characterization of heterotrophic nitrification and aerobic denitrification by Alcaligenes faecalis NS13)

  • 정택경;라창식;조기성;송홍규
    • 미생물학회지
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    • 제52권2호
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    • pp.166-174
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    • 2016
  • 호기적 조건에서 질산화와 탈질화를 동시에 진행하는 Alcaligenes faecalis NS13 균주를 분리하여 다양한 특성을 파악하였다. 이 균주는 $15-37^{\circ}C$ 온도에서 생장할 수 있으며 암모니움 산화율이 높고 고농도의 암모니움 환경에서도 생장이 저해되지 않고 초기 암모니움 농도 증가에 따라 제거량이 증가하였다. pH와 염분농도에 대해서도 내성 범위가 넓어 암모니움 산화가 영향을 받지 않았다. 질산화에 이어진 탈질화로 인해 질산염의 축적이 일어나지 않았으며 탈질화의 중간산물인 아산화질소는 미량 검출되었지만 배양 후 모든 질소 화합물을 측정한 결과 약 42.8%가 $N_2$로 전환된 것으로 추정되었다. 탈질화는 PCR 증폭을 통해서 탈질화에 관여하는 유전자 nitrate reductase gene, napA과 nitrous oxide reductase gene, nosZ의 존재로 뒷받침되었다. 또한 배지 내 질소의 46.4%가 NS13 균주로 동화되었기 때문에 폐수처리 시 질산화 및 탈질화 후에 슬러지로 처분한다면 실질적으로 89% 이상의 우수한 암모니움의 제거효과를 거둘 수 있을 것이다.

Isolation and Characterization of Thioredoxin cDNA from Codonopsis lanceolata (S. et Z.) Trautv

  • In, Jun-Gyo;Lee, Bum-Soo;Rho, Yeong-Deok;Yu, Chang-Yeon;Yang, Deok-Chun
    • 한국약용작물학회지
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    • 제13권5호
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    • pp.293-297
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    • 2005
  • A thoredoxin (CTRX) gene was cloned and characterized from a full length cDNA library prepared from taproot of three-year old Codonopsis lanceolata. A CTRX was 666 nucleotides long and has an open reading frame of 372 bp with 124 amino acid residues (pI = 4.92). The deduced amino acid sequence of the CTRX matched to the previously reported plant thioredoxin h genes. The deduced amino acid sequence of CTRX exhibited the similarity of 33-67% among previously registered thioredoxin genes. The expression of CTRX in leaves of Codonopsis lanceolata was increased by wounding and 1 mM $H_2O_2$, but decreased by 0.1 mM cadmium.

Gene Microarray Analysis for Porcine Adipose Tissue: Comparison of Gene Expression between Chinese Xiang Pig and Large White

  • Guo, W.;Wang, S.H.;Cao, H.J.;Xu, K.;Zhang, J.;Du, Z.L.;Lu, W.;Feng, J.D.;Li, N.;Wu, C.H.;Zhang, L.
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권1호
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    • pp.11-18
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    • 2008
  • We created a cDNA microarray representing approximately 3,500 pig genes for functional genomic studies. The array elements were selected from 6,494 cDNA clones identified in a large-scale expressed sequence tag (EST) project. These cDNA clones came from normalized and subtracted porcine adipose tissue cDNA libraries. Sequence similarity searches of the 3,426 ESTs represented on the array using BLASTN identified 2,790 (81.4%) as putative human orthologs, with the remainder consisting of "novel" genes or highly divergent orthologs. We used the gene microarray to profile transcripts expressed by adipose tissue of fatty Chinese Xiang pig (XP) and muscley Large White (LW). Microarray analysis of RNA extracted from adipose tissue of fatty XP and muscley LW identified 81 genes that were differently expressed two fold or more. Transcriptional differences of four of these genes, adipocyte fatty acid binding protein (aP2), stearyl-CoA desaturase (SCD), sterol regulatory element binding transcription factor 1 (SREBF1) and lipoprotein lipase (LPL) were confirmed using SYBR Green quantitative RT-PCR technology. Our results showed that high expression of SCD and SREBF1 may be one of the reasons that larger fat deposits are observed in the XP. In addition, our findings also illustrate the potential power of microarrays for understanding the molecular mechanisms of porcine development, disease resistance, nutrition, fertility and production traits.

Association between PCR-RFLP Polymorphism of the Fifth Intron in Lipoprotein Lipase Gene and Productive Traits in Pig Resource Family

  • Zhang, B.Z.;Lei, M.G.;Deng, C.Y.;Xiong, Y.H.;Zuo, B.;Li, F.E.
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권4호
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    • pp.458-462
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    • 2005
  • The study was aimed at detecting polymorphism of the fifth intron in lipoprotein lipase (LPL) gene and analyzing association between the polymorphism and productive traits. A pair of primers was designed for amplifying the fifth intron. Sequence analysis indicated that a G1171C substitution existed in Large White breed. The mutation was detected by PCR-AfaI-RFLP. Polymorphism analysis in a pig resource family showed that there existed significant effects on carcass and meat quality traits. Thoraxwaist fat thickness of BB genotype was significantly higher (14.2%, p<0.05) than that of AA on carcass traits, while BB genotype was significantly lower (3.6% p<0.01, 4.1% p<0.01; 2.3% p<0.01, 1.9% p<0.01; 1.8% p<0.01, 1.4% p<0.05) than AA and AB genotype in pH of m. Longissimus Dorsi (LD), m. Biceps Femoris (BF), m. Semipinali Capitis (SC). The allelic frequencies were also significantly different between indigenous Chinese breeds and exotic breeds. Data analyzed revealed that the mutation locus affected production traits mostly by additive effects. Based on these results, it is necessary to do more studies on LPL gene before making the LPL locus into the application of marker-assisted selection (MAS) programs.

ASCL2 Gene Expression Analysis and Its Association with Carcass Traits in Pigs

  • Cheng, H.C.;Zhang, F.W.;Deng, C.Y.;Jiang, C.D.;Xiong, Y.Z.;Li, F.E.;Lei, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권10호
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    • pp.1485-1489
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    • 2007
  • Achaete-scute like 2 (ASCL2) gene encodes a member of the basic helix-loop-helix transcription factor which is essential for the maintenance of proliferating trophoblasts during placental development. ASCL2 gene preferentially expresses the maternal allele in the mouse. However, it escapes genomic imprinting in the human. In this study, the complete open reading frame consisting of 193 amino acids of ASCL2 gene was obtained. Sequence analysis indicated that a C-G mutation existed in the 3' region between Meishan and Large White pigs. The polymorphism was used to determine the monoallelic or biallelic expression with RT-PCR-RFLP in pigs of Large $White{\times}Meishan$ $F_1$ hybrids. Imprinting analysis indicated that the ASCL2 gene expression was biallelic in all the tested tissues (heart, liver, spleen, lung, kidney, stomach, small intestine, skeletal muscle, fat, uterus, ovary and pituitary). PCR-RFLP was used to detect the polymorphism in 270 pigs of the "$Large\;White{\times}Meishan$" $F_2$ resource population. The statistical results showed highly significant associations of the genotypes and fat meat percentage (FMP), lean meat percentage (LMP) and ratio of lean to fat (RLF) (p<0.01), and significant associations of the genotypes and loin eye area (LEA) and internal fat rate (IFR) (p<0.05).

Expression and Characterization of the Human Lactoferrin in the Milk of Transgenic Mice

  • Z. Y. Zheng;Y. M. Han;Lee, K. K.
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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    • pp.85-85
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    • 2003
  • Human lactoferrin (hLF) is an 80 kDa iron-binding glycoprotein that is expressed in high concentration in milk and in lesser amount in the secondary or specific granules of neutrophils and in plasma, LF is classically considered to be related to the binding, transport, and storage of iron. The transgenic mice carrying the human hLF gene in conjunction with the bovine $\beta$-casein promoter produced the human hLF in their milk during lactation. To screen transgenic mice, PCR was carried out using chromosomal DNA extracted from tail or toe tissues. In this study, stability of germ line transmission and expression of hLF were monitored up to generation Fl7 of a transgenic line. When female mouse of generation F9 was crossbred with normal male, generation F9 to Fl7 mice showed similar transmission rates ($66.0 \pm 12.57%, 42.0 \pm 14.98%, 72.2 \pm 25.45%, 50.0 \pm 16.70%, 65.7 \pm 6.45%, 48.6 \pm 14.65%, 54 1 \pm 18 11%, 57.8 \pm 16.16% and 48.6 \pm 20.66$, respectively), implying that the hLF gene can be transmitted stably up to long term generation in the transgenic mice For ELISA analysis, hLF expression levels were determined with an hLF ELISA kit in accordance with the supplier's protocol. Expression levels of human hLF from milk of generation F9 to Fl3 mice were $ 3.2 \pm 0.69 mg/ml, 3.1 \pm 0.81 mg/ml, 4.6 \pm 1.38 mg/ml, 3.1 \pm 0.42 mg/ml, and 4.5 \pm 1,48 mg/ml$, respectively. These expression levels were lower than that of founder (6.6 mg/$m\ell$) mouse. We concluded that transgenic mice faithfully passed the transgene on their progeny and successively secreted target proteins into their milk through several generations.

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Association between PCR-RFLP Polymorphisms of Five Gene Loci and Milk Traits in Chinese Holstein

  • Zhang, R.F.;Chen, H.;Lei, C.Z.;Fang, X.T.;Zhang, Y.D.;Hu, S.R.;Su, L.H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권2호
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    • pp.166-171
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    • 2007
  • The objective of this study was to assess the association of polymorphisms in ${\kappa}$-cn, ${\beta}$-lg, ${\beta}$-lg 5′ flanking region, CSN1S2, and IGFBP-3 genes with milk production traits and mastitis-related traits in Chinese Holstein. Traits analyzed were 305 day standard milk yield, protein percentage, fat percentage, the ratio of fat percentage and protein percentage, pre-somatic cell count, somatic cell count, and somatic cell score, respectively. CSN1S2 locus was uninformative because only one genotype BB was found in Chinese Holstein. Allele frequencies of A and B in IGFBP-3 gene were 0.5738 and 0.4262 in Chinese Holstein population, which was different from reported Qinchuan cattle population. The genotypes of animals at IGFBP-3 locus significantly affected 305 day standard milk yield, protein percentage, and somatic cell score. The ${\beta}$-lg genotypes had a significant effect on protein percentage and the ratio of fat percentage and protein percentage. Polymorphism in ${\beta}$-lg 5′ flanking region was associated with 305 day standard milk yield, protein percentage, fat percentage, pre-somatic cell count, and somatic cell count. No significant associations of the polymorphism in ${\kappa}$-cn gene were observed for any trait.

Estimation of Gene Effects in Four Bivoltine Silkworm (Bombyx mori L.) Crosses

  • Malik, G.N.;Singh, T.P.;Rufaie, S.Z.Haque;Aijaz, M.;Dar, H.U.
    • International Journal of Industrial Entomology and Biomaterials
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    • 제8권1호
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    • pp.113-115
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    • 2004
  • Six generations (P$_1$, P$_2$, F$_1$, BC$_1$, BC$_2$ and F$_2$) of four bivoltine silkworm crosses (SKAU-R-1 ${\times}$ Yakwei, SKAU-R-6 ${\times}$ Yakwei, CSR$_2$ ${\times}$ CSR$_4$ and SH$_{6}$${\times}$ NB$_4$D$_2$) were evaluated in a completely randomized block design with 5 replications for each treatment. The generation mean 2 in respect of 3 metric traits (single cocoon weight, single shell weight, and shell ratio %), were subjected to Cavallis joint scaling test. Additive dominance model was found to be adequate in CSR$_2$${\times}$CSR$_4$ and SH$_{6}$${\times}$ NB$_4$D$_2$ for single cocoon weight and shell weight and SKAU-R-6 ${\times}$ Yakwei for shell ratio(%). Whereas, in rest of the crosses epistasis was evident in the traits under investigation. Magnitude of additive gene effect (d) was greater than dominance(h) in SH$_{6}$${\times}$NB$_4$D$_2$ and SKUA-R-6${\times}$Yakwei for shell ratio (%) and in CSR$_2$${\times}$ CSR$_4$ for shell weight. Thus selection for these traits in early segregating generations of these crosses would be effective for obtaining considerable genetic gain. gain.

Effect of Cellular Zinc on the Regulation of C2-ceramide Induced Apoptosis in Mammary Epithelial and Macrophage Cell Lines

  • Han, S.E.;Lee, H.G.;Yun, C.H.;Hong, Z.S.;Kim, S.H.;Kang, S.K.;Kim, S.H.;Cho, J.S.;Ha, S.H.;Choi, YunJaie
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권12호
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    • pp.1741-1745
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    • 2005
  • Zinc is a trace element that is associated with a stimulation of immune function and regulation of ion balance for livestock production. In this study, the effect of zinc as inhibitor to apoptosis-induced cells was examined in vitro using mammary epithelial cell line, HC11 and macrophage cell line, NCTC3749. Cell viability, measured by MTT assay, indicated that 10 g/ml of zinc had a negative impact on cellular activity and 50 ng/ml was chosen for further testing. Apoptosis was induced in cells treated with C2-ceramide in serum-free media. DNA fragmentation and gene expression of acidic sphingomyelinase (a gene responsible for the progress of apoptosis) were distinctively low in zinc treated cells compared with those in non-treated controls. In conclusion, zinc is involved in the regulation of cell proliferation and apoptosis in mammary epithelial cells and macrophages.