• Title/Summary/Keyword: lacZ/lacI

Search Result 47, Processing Time 0.03 seconds

Transfer of Foreign Gene into Mud Loach, Misgurnus mizolepis I . Availability of the lacZ as a reporter gene for producing transgenic mud loach (미꾸라지, Misgurnus mizozepis에 외래 유전자 이식 I. lacZ의 reporter 유전자로서의 유용성 검토)

  • KIM Dong Soo;NAM Yoon Kwon
    • Journal of Aquaculture
    • /
    • v.7 no.1
    • /
    • pp.41-54
    • /
    • 1994
  • In order to evaluate the availability of lacZ as a reporter gene for producing transgenic mud loach, foreign DNA, bacterial \beta-galactosidase$ gene (lacZ) was microinjected into mud loach eggs and its insertion and expression were examined. X-gal based histochemical assay, fluorimetric analysis of \beta-galactosidase$ with 4-methylumbelliferyl-$\beta$-D-galactoside (MUG) and molecular biological examination using polymerase chain reaction (PCR), dot blot, southern blot and sequence analysis of PCR products were carried out to analyze both microinjected group and non-injected controls. The results are disccussed in this paper.

  • PDF

Use of Transgenic and Mutant Animal Models in the Study of Heterocyclic Amine-induced Mutagenesis and Carcinogenesis

  • Dashwood, Roderick H.
    • BMB Reports
    • /
    • v.36 no.1
    • /
    • pp.35-42
    • /
    • 2003
  • Heterocyclic amines (HCAs) are potent mutagens generated during the cooking of meat and fish, and several of these compounds produce tumors in conventional experimental animals. During the past 5 years or so, HCAs have been tested in a number of novel in vivo murine models, including the following: lacZ, lacI, cII, c-myc/lacZ, rpsL, and $gpt{\Delta}$ transgenics, $XPA^{-/-}$, $XPC^{-/-}$, $Msh2^{+/-}$, $Msh2^{-/-}$ and $p53^{+/-}$ knock-outs, Apc mutant mice ($Apc^{{\Delta}716}$, $Apc^{1638N}$, $Apc^{min}$), and $A33^{{\Delta}N{\beta}-cat}$ knock-in mice. Several of these models have provided insights into the mutation spectra induced in vivo by HCAs in target and non-target organs for tumorigenesis, as well as demonstrating enhanced susceptibility to HCA-induced tumors and preneoplastic lesions. This review describes several of the more recent reports in which novel animal models were used to examine HCA-induced mutagenesis and carcinogenesis in vivo, including a number of studies which assessed the inhibitory activities of chemopreventive agents such as 1,2-dithiole-3-thione, conjugated linoleic acids, tea, curcumin, chlorophyllin-chitosan, and sulindac.

Construction of an Escherichia-Pseudomonas Shuttle Vector Containing an Aminoglycoside Phosphotransferase Gene and a lacZ' Gene for $\alpha$-Complementation

  • Lee, Bheong-Uk;Hong, Ja-Heon;Kahng, Hyung-Yeel;Oh, Kye-Heon
    • Journal of Microbiology
    • /
    • v.44 no.6
    • /
    • pp.671-673
    • /
    • 2006
  • A new 4.87 kb Escherichia-Pseudomonas shuttle vector has been constructed by inserting a 1.27 kb DNA fragment with a replication origin of a Pseudomonas plasmid pRO1614 into the 3.6 kb E. coli plasmid pBGS18. This vector, designated pJH1, contains an aminogly-coside phosphotransferase gene (aph) from Tn903, a lacZ' gene for $\alpha$-complementation and a versatile multiple cloning site possessing unique restriction sites for EcoRI, SacI, KpnI, SmaI, BamHI, XbaI, SalI, BspMI, PstI, SphI, and HindIII. When pJH1 was transformed into E. coli DHS${\alpha}$ and into P. putida HK-6, it was episomally and stably maintained in both strains. In addition, the enhanced green fluorescent protein (EGFP) gene which was transcriptionally cloned into pJH1 rendered E. coli cells fluorescence when its transformants were illuminated at 488 nm.

Isolation of Lactococcus lactis Strain with ${\beta}$-Galactosidase Activity from Kimchi and Cloning of lacZ Gene from the Isolated Strain

  • Park, Rae-Jun;Lee, Kwang-Hee;Kim, Su-Jung;Park, Jae-Yong;Nam, Su-Jin;Yun, Han-Dae;Lee, Hyong-Joo;Chang, Hae-Choon;Chung, Dae-Kyun;Lee, Jong-Hoon;Park, Yun-Hee;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
    • /
    • v.12 no.1
    • /
    • pp.157-161
    • /
    • 2002
  • A lactic acid bacteria with ${\beta}$-gal activity was isolated from Kimchi, a traditional fermented vegetable food in Korea. The isolate was identified as a Lactococcus lactis strain and named L. lactis A2. The gene encoding ${\beta}$-gal of L. lactis A2 was cloned as a 5.8 kb PstI fragment. DNA sequencing identified the complete lacA (galactoside acetyltransferase)-lacZ (${\beta}$-galactosidase) genes together with the 3' part of upstream galT (galactose-1-phosphate uridyltransferase), and the 5'region of downstream galE (UDP-galactose-4-epimerase) genes. L. lactis A2 had the same gal/lac operon structure as in L. lactis subsp. lactis 7962. Other genes of the Leloir pathway are most likely to be located in the 5'upstream of the 5.8 kb fragment on the A2 chromosome. Sequences downstream of galE were different from those of L. lactis subsp. lactis 7962.

The influence of MgO on the radiation protection and mechanical properties of tellurite glasses

  • Hanfi, M.Y.;Sayyed, M.I.;Lacomme, E.;Akkurt, I.;Mahmoud, K.A.
    • Nuclear Engineering and Technology
    • /
    • v.53 no.6
    • /
    • pp.2000-2010
    • /
    • 2021
  • Mechanical moduli, such as Young's modulus (E), Bulks modulus (B), Shear modulus (S), longitudinal modulus (L), Poisson's ratio (σ) and micro Hardness (H) were theoretically calculated for (100-x)TeO2+x MgO glasses, where x = 10, 20, 30, 40 and 45 mol%, based on the Makishima-Mackenzie model. The estimated results showed that the mechanical moduli and the microhardness of the glasses were improved with the increase of the MgO contents in the TM glasses, while Poisson's ratio decreased with the increase in MgO content. Moreover, the radiation shielding capacity was evaluated for the studied TM glasses. Thus, the linear attenuation coefficient (LAC), mass attenuation coefficient (MAC), transmission factor (TF) and half-value thickness (𝚫0.5) were simulated for gamma photon energies between 0.344 and 1.406 MeV. The simulated results showed that glass TM10 with 10 mol % MgO possess the highest LAC and varied in the range between 0.259 and 0.711 cm-1, while TM45 glass with 45 mol % MgO possess the lowest LAC and vary in the range between 0.223 and 0.587 cm-1 at gamma photon energies between 0.344 and 1.406 MeV. Furthermore, the BXCOM program was applied to calculate the effective atomic number (Zeff), equivalent atomic number (Zeq) and buildup factors (EBF and EABF) of the glasses. The effective removal cross-section for the fast neutrons (ERCSFN, ∑R) was also calculated theoretically. The received data depicts that the lowest ∑R was achieved for TM10 glasses, where ∑R = 0.0193 cm2 g-1, while TM45 possesses the highest ERCSFN where ∑R = 0.0215 cm2 g-1.

Incapability of Utilizing Galactose by pgs1 Mutation Occurred on the Galactose Incorporation Step in Saccharomyces cerevisiae

  • Rho, Min-Suk;Su, Xuefeng;Lee, Yoon-Shik;Kim, Woo-Ho;Dowhan, William
    • Journal of Microbiology and Biotechnology
    • /
    • v.16 no.1
    • /
    • pp.84-91
    • /
    • 2006
  • A Saccharomyces cerevisiae pgs1 nulI mutant, which is deficient with phosphatidyl glycerol (PG) and cardiolipin (CL) biosynthesis, grows well on most fermentable carbon sources, but fails to grow on non-fermentable carbon sources such as glycerol, ethanol, and lactate. This mutant also cannot grow on galactose medium as the sole carbon source. We found that the incorporation of $[^{14}C]-galactose$, which is the first step of the galactose metabolic pathway (Leloir pathway), into the pgs 1 null mutant cell was extremely repressed. Exogenously expressed PGS1 (YCpPGS1) under indigenous promoter could completely restore the pgs1 growth defect on non-fermentable carbon sources, and dramatically recovered $[^{14}C]-galactose$ incorporation into the pgs1 mutant cell. However, PGS1 expression under the GALl promoter $(YEpP_{GAL1}-PGS1myc)$ could not complement pgs1 mutation, and the GAL2-lacZ fusion gene $(YEpP_{GAL2}-lacZ)$ also did not exhibit its $\beta-galactosidase$ activity in the pgs1 mutant. In wild-type yeast, antimycin $A(1\;{\mu}g/ml)$, which inhibits mitochondrial complex III, severely repressed not only the expression of the GAL2-lacZ fusion gene, but also uptake of $[^{14}C]-galactose$. However, exogenously expressed PGS1 partially relieved these inhibitory effects of antimycin A in both the pgs1 mutant and wild-type yeast, although it could not basically restore the growth defect on galactose by antimycin A. These results suggest that the PGSI gene product has an important role in utilization of galactose by Gal genes, and that intact mitochondrial function with PGS1 should be required for galactose incorporation into the Leloir pathway. The PGS1 gene might provide a clue to resolve the historic issue about the incapability of galactose with deteriorated mitochondrial function.

The Schizosaccharomyces pombe Gene Encoding ${gamma}-Glutamyl$ Transpeptidase I Is Regulated by Non-fermentable Carbon Sources and Nitrogen Starvation

  • Kim, Hong-Gyun;Park, Hey-Jung;Kang, Hyun-Jung;Lim, Hye-Won;Kim, Kyung-Hoon;Park, Eun-Hee;Ahn, Ki-Sup;Lim, Chang-Jin
    • Journal of Microbiology
    • /
    • v.43 no.1
    • /
    • pp.44-48
    • /
    • 2005
  • In our previous study, the first structural gene (GGTI) encoding ${\gamma}-glutamyl$ transpeptidase was cloned and characterized from the fission yeast Schizosaccharomyces pombe, and its transcription, using the GGTI-lacZ fusion gene, containing the 1,085 bp upstream region from the translational initiation point, was found to be enhanced by sodium nitroprusside and L-buthionine-(S,R)-sulfoximine (BSO). In the present work, regulation of the GGTI gene was further elucidated. Non-fermentable carbon sources, such as acetate and ethanol, markedly enhanced the synthesis of ${beta}-galactosidase$ from the GGTI-lacZ fusion gene. However, its induction by non-fermentable carbon sources appeared to be independent of the presence of the Pap1 protein. Nitrogen starvation also gave rise to induction of GGTI gene expression in a Pap1-independent manner. The three additional fusion plasmids, carrying 754, 421 and 156 bp regions, were constructed. The sequence responsible for the induction by non-fermentable carbon sources and nitrogen starvation was identified to exist within a -421 bp region of the GGTI gene. Taken together, the S. pombe GGTI gene is regulated by non-fermentable carbon sources and nitrogen starvation.

Regulation of stf Operon Expression and Its Virulence (살모넬라가 발현하는 stf 오페론의 조절과 병원성 인자로서의 기능)

  • Kim Sam-Woong;Kim Young-Hee;Kang Ho-Young
    • Journal of Life Science
    • /
    • v.15 no.4 s.71
    • /
    • pp.553-560
    • /
    • 2005
  • The stf (Salmonella typhimurium fimbriae) operon consisting of stfA(CDEFG assumes to encode putative fimbriae. The complete stf operon is existed in S. typhimurium and S. choleraesuis, whereas it is absent in S. typhi. Analyses of the amino acid residues between major subunit StfA of the Stf fimbriae and those of known other fimbriaes suggested that Stf belongs to class I type fimbriae. Through comparison of StfD chaperone with the other fimbrial chaperones, and of C-terminus in subunits of Stf fimbriae, it belongs to FGS (with a short Fl-G1 loop) subfamily. In order to investigate the expression of stf operon, we have constructed a Salmonella strain containing a chromosomal stfA::lacZYA transcriptional fusion, resulting in S. typhimurium $_X8532$. The strain $_X8532$ lacked the expression of \beta-galactosidase$ under normal culture conditions. However, with longer incubation time of the S. typhimurium $_X8532$, we have isolated 21 individual strains exhibiting $Lac^+$ phenotype. $Lac^+$ phenotype was appeared as approximately 0.03 frequency per generation. All isolates expressed lacZ constitutively in the various environmental conditions. Various global regulatory proteins including RpoS, OmpR, and CpxR were not involved in the regulation of the stf operon. A S. typhimurium $_X8661$ mutant lacking stfAC function attenuated 6.7 folds more than that of wild type $_X3761$ in the mouse virulence test, suggesting in the somehow involved in the Salmonella pathogenesis.

Expression of mue Gene on Plasmid pKM101 and pSL4 (플라스미드 pKM101 과 pSL4 의 muc 유전자의 발현에 관한 연구)

  • 전홍기;황유경;이상률;백형석
    • Korean Journal of Microbiology
    • /
    • v.30 no.5
    • /
    • pp.371-376
    • /
    • 1992
  • Plasmid pSL4 of plasmid pKM 101 mutant have high protection effects and mutagenecity for UV and methyl methanesulfonate, The mucA gene and a pan of mucE gene of pKM 101 and pSL4 were sucloned onto lacZ' fusion vector pMC874 and the hybrid plasmids pBH31 and pBH30 were selected. These plsmids were intrduced into $recA^{+}lexA^{-}$, $recA^{-}와lexA^{+}$ strains and determined the activity of $\beta$-galactosidase for UV. In $recA^{+}lexA^{+}$ strain.$\beta$-galactosidase activity of pBH30 included mue region of pSL4 was higher thall pBH31 inclued muc region of pKM 10 I and the tf-galactosidase of two plasmids was not induced in reeA and leeA mutants with or without UV illumination. Without UV illumination. the .$\beta$-galactosidasc of pBH30 was expressed a little higher level than that of pBH3L We suggest that the functional difference of pKM 10l and pSL4 are due to the variety of mue regulatory region. Also. a plasmid pBH 100 earring umuC' -lacZ' gene fusion was constructed in vitro to study the regulation of the umu operon. It was shown that the umu operon is induced by UV and is regulated by the reeA and lexA genes.

  • PDF

Zygotic Expression of c-myc Gene in Mouse Early Embryos: Functional Role of c-myc Promoter (생쥐 초기배아에서 c-myc Proto-Oncogene Promoter의 기능적 활성화)

  • Park, Ki-Soo;Kang, Hae-Mook;Shim, Chan-seob;Sun, Woong;Kim, Jae-man;Lee, Young-Ki;Kim, Kyung-jin
    • The Korean Journal of Zoology
    • /
    • v.38 no.4
    • /
    • pp.550-556
    • /
    • 1995
  • The c-myc proto-oncogene is Involved In the control of normal cell proliferation and differentiation of many cell lineages. Although it has heen suggested that c-myc may play an important role in the mammalian early development, it Is unclear whether the embryonic c-myc mRNA is originated from zygotic gene expression or stored maternal message. Thus, we have construded expression vectors, In which the 5, flanking sequences including c-myc promoter region and a large non-coding exon I are fused 'sith E. coli lacZ gene that encedes $\beta$-galactosldase as a reporter. As c-myc exon I contains a modulatory sequence, we designed t, vo types of vectors (pcmyc.Gall and pcmyc-Ga12) to examine the role of exon I in c-myc expression. The former contains the complete exon I and the later has a deletion in 40 bp of modulator sequence located In the exon I of c-myc These vectors were microInjected into fertilized one-cell embryos and $\beta$-galactosidase activity was examined by X-gal staining during early embryogenesis. $\beta$-galactosidase activity derived from c-myc promoter was decreased at two-cell stage. The expression level directed by pcmyc- Ga12 was similar to that of pcmyc-Gal1, indicating that the medulatory sequence in exon I may not be Involved at least In the regulation of embryonic c-myc expression. In summary, the present study indicates that the c-myc promoter is functional at the early stage embryo, and the regulation of c-myc expression is under the control of "zygotic" clock of preimplantation mouse embryos.e embryos.

  • PDF