• Journal of Microbiology and Biotechnology
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• v.16 no.2
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• pp.286-290
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• 2006

A genomic library of Streptococcus vestibularis ATCC49124 was constructed in an E. coli plasmid vector, and the urease-positive transformants harboring the urease gene cluster were isolated on Christensen-urea agar plates. The minimal DNA region required for urease activity was located in a 5.6 kb DNA fragment, and a DNA sequence analysis revealed the presence of a partial ureI gene and seven complete open reading frames, corresponding to ureA, B, C, E, F, G, and D, respectively. The nucleotide sequence over the entire ure gene cluster and 3'-end flanking region of S. vestibularis was up to 95% identical to that of S. salivarius, another closely related oral bacterium, and S. thermophilus, isolated from dairy products. The predicted amino acid sequences for the structural peptides were 98-100% identical to the corresponding peptides in S. salivarius and S. thermophilus, respectively, whereas those for the accessory proteins were 96-100% identical. The recombinant E. coli strain containing the S. vestibularis ure gene cluster expressed a high level of the functional urease holoenzyme when grown in a medium supplemented with 1 mM nickel chloride. The enzyme was purified over 49-fold by using DEAE-Sepharose FF, Superdex HR 200, and Mono-Q HR 5/5 column chromatography. The specific activity of the purified enzyme was 2,019 U/mg, and the Michaelis constant ($K_{m}$) of the enzyme was estimated to be 1.4 mM urea. A Superose 6HR gel filtration chromatography study demonstrated that the native molecular weight was about 196 kDa.

• Fisheries and Aquatic Sciences
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• v.15 no.1
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• pp.43-47
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• 2012

Growth hormone (GH) transgenesis in fish has the potential to improve aquaculture efficiency and capacity. However, many fast-growing transgenic fish have experienced side effects caused by excess GH expression. To overcome this unwanted issue associated with several GH transgenic mud loach Misgurnus mizolepis lines carrying GH construct driven by a strong ${\beta}$-actin regulator ($pml{\beta}$-actGH), we performed an alternative version of GH autotransgenesis using a weaker but more stable regulator, the mud loach lectin promoter. GH transgenesis with a pmlectGH construct consisting of the mud loach GH gene driven by the 2.3-kb lectin promoter exhibited significant growth stimulation. However, the extent of the growth acceleration in pmlectGH transgenics (six times maximum when assessed 2 months post hatching) was much less than that in transgenic individuals carrying the $pml{\beta}$-actGH construct. Additionally, the extraordinary gigantism that was common in $pml{\beta}$-actGH-transgenic mud loaches was diminished in transgenic loaches harboring the pmlectGH construct. Transgenic founders (pmlectGH) successfully transmitted their transgene into the next generation with up to 41% frequency. Growth stimulation also persisted in the transgenic F1 strains, with a seven-fold increase in maximum body weight at 6 months of age.

• Journal of Embryo Transfer
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• v.21 no.1
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• pp.13-20
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• 2006

The purpose of this study was to develope the transgenic cattle expressing hFSH into the urine using the nuclear transfer. To produce the interest gene in urine, the specific vector was ligated with hFSH gene undo. maUII promoter. The fetal fibroblast cells (KbFF) were isolated from a 45-day male fetus. The hFSH gene was co-transfected with pcDNA3 (neo) vector to KbFF cells by electroporation. The gene-transfected cells were cultured with G-418 selection medium for 2 weeks. Selected colonies were confirmed by PCR. For nuclear transfer, enucleated bovine oocytes were transferred with hFSH transfected or nontransfected fetal fibroblasts. The cleavage and blastocyst formation rates were significantly lower (p<0.05) in cloned embryos transfected with hFSH gene (68.7% and 15.7%) than in those non-transfected (67.6% and 24.5 %), respectively. Apoptosis analysis showed no difference between hFSH transfected and non-transfected blastocysts (p>0.05). The blastocysts were transfected to 77 (control 24, hFSH 53) recipient cows. Two calves were born (1.9%) following transfer with NT embryos transfected with hFSH gene, but they were confirmed not to be transgenic calves. This result shows that the hFSH colonies were mixed with transfected and non transfected cells. Further research will be needed for selection and establishment of gene transfected cells.

• Mycobiology
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• v.37 no.4
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• pp.258-266
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• 2009

Pleurotus eryngii, known as king oyster mushroom has been widely used for nutritional and medicinal purposes. This study was initiated to screen the suitable conditions for mycelial growth and to determine the phylogenetic relationship of the selected strains. Optimal mycelial growth was observed at $30{^{\circ}C}$ and minimum mycelial growth observed at $10{^{\circ}C}$. This mushroom tolerates a broad pH range for mycelial growth, with most favorable growth observed at pH 6. Results also indicated that glucose peptone, yeast malt extract and mushroom complete media were favorable growth media, while Hennerberg and Hoppkins media were unfavorable. Dextrin was the best and xylose the least effective carbon sources. Results revealed that inorganic nitrogen sources were less effective than organic sources for the mycelial growth of P. eryngii. Investigation of genetic diversity is necessary to identify the strains. The ITS region of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 214 to 222 bp and 145 to 236 bp, respectively. The sequence of ITS2 was more variable than that of ITS1, and the 5.8S sequences were identical. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into six clusters. Fourteen IUM and ATCC- 90212 strains were also analyzed by RAPD with 20 arbitrary primers. Fourteen of these primers were efficiently amplified the genomic DNA. The number of amplified bands varied with the primers and strains, with polymorphic fragments in the range from 0.2 to 2.3 kb.

• Economic and Environmental Geology
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• v.18 no.2
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• pp.139-150
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• 1985

Skarn at the Dielette formed largely in calc-silicate hornfels at the contact with the Flamanville granite. The skarn consists mainly of garnet and pyroxene, and less frequently vesuvianite. Traversing toward calc-silicate hornfels wall rock from a central zone of the skarn, the general sequence of formation of mineral assemblages is: (1) dark brown garnet (2) pale brown garnet-vesuvianite-pyroxene, and (3) pyroxene-prehnite-scapolite-wollastonite envelopes (designated as transition zone) developed between skarn and calc-silicate hornfels. The central zone of the skarn consists mainly of dark brown garnets (garnet I) that contain little or no pyroxene. The pale brown garnet (garnet II) is associated with pyroxene and vesuvianite. The sequence of these garnets results from the zonal growth outward. There is an abrupt discontinuity in composition between garnet I formed in early stage and garnet II in late stage, while each garnet shows relatively uniform composition. At the zone in contact with the granite, the iron contents of garnets decrease toward the marginal zone of the skarn, from an average value of 36 mole % andradite in garnet I to 18 mole % andradite in garnet II. At the zone distant from the granite, the andradite component decreases from 28 mole % in garnet 1 to 19 mole % in garnet II. The variation of the iron contents of pyroxenes is also similar to that of garnets. The sharp discontinuity in composition of garnets and pyroxenes suggests that the skarn of study area was formed by infiltration metasomatic process. The results of the analyses of mineral assemblages of the transition zone by chemical potential diagrams suggest that the transition zone was made by the diffusion of the elements Ca, K and Fe from the skarn to the calc-silicate hornfels contact zone. The estimated temperatures and $Xco_2$ for the formation of the transition zone show $300^{\circ}C$$440^{\circ}C$ and $0.07{\pm}0.05<Xco_2<0.02{\pm}0.01$ at 1 Kb respectively.

• Economic and Environmental Geology
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• v.8 no.1
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• pp.1-23
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• 1975

The subject of this study deals with phase relations between stannite ($Cu_2FeSnS_4$) and sphalerite (${\beta}-ZnS$)/wurtzite (${\alpha}-ZnS$). The phase relations were systematically investigated from liquidus temperature to $400^{\circ}C$ under controlled conditions. ${\beta}-stannite$ (tetragonal) is stable up to $706{\pm}5^{\circ}C$, where it inverts to a high-temperature polymorph ${\alpha}-stannite$ (cubic) melting congruently at $867{\pm}5^{\circ}C$. Sphalerite (cubic, ${\beta}-ZnS$) inverts at $1013{\pm}3^{\circ}C$ to wurtzite, which is the hexagonal hightemperature polymorph of ZnS. Between ${\alpha}-stannite$ and sphalerite a complete solid solution series exists above approximately $870^{\circ}C$ up to solidus temperature. The melting temperature of ${\alpha}-stannite$ rises towards sphalerite and reaches a maximum at $1074{\pm}3^{\circ}C$, which is the peritectic with the composition of 91 wt. % sphalerite and 9 wt. % ${\alpha}-stannite$. At this temperature, wurtzite takes only 5wt. % ${\alpha}-stannite$ in solid solution which decreases with increasing temperature. The inverson temperature of ${\alpha}/{\beta}-stannite$ is lowered with increasing amounts of sphalerite in solid solution down to $614{\pm}7^{\circ}C$, which is the eutectoid with the composition of 13 wt. % sphalerite and 87 wt. % ${\alpha}-stannite$. Here, ${\beta}-stannite$ contains only 10wt. % sphalerite in solid solution. With decreasing temperature, the ranges of the solid solution on both sides of the system narrow. The phase relations in the above pure system changed due to the FeS impurities in the sphalerite solid solution. The eutectoid increased from $614{\pm}7^{\circ}C$ up to $695{\pm}5^{\circ}C$ (5 wt. % FeS) and $700{\pm}5^{\circ}C$ (10wt. % FeS), while the peritectic decreased from $1074{\pm}3^{\circ}C$ down to $1036{\pm}3^{\circ}C$ (wt. %FeS) and $987{\pm}3^{\circ}C$ (10wt. %FeS). A most notable change is the appearance of non-binary regions. An important feature is the combination of this study system with the experimental results reported by Sprinfer (1972). If a stannite-kesterite solid solution is used in the place of stannite as a bulk composition, the inversion temperature is lowered to less than $400^{\circ}C$ which belongs to temperatures of the hydrothermal region.

• Journal of dental hygiene science
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• v.3 no.1
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• pp.11-14
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• 2003

A methylotrophic Actinomycetes strain, which produce the anti-oral cancer activity compound, was isolated from soil and estimated as Amylocolatosis sp. based on taxonomic studies. A methanol didn't have influence on the production of the anticancer compounds. These compound were isolated by ethylacetate extract, silica gel column chromatography, sephadex LH-20 column and reverse phase HPLC. The compounds were very stable under heat ($121^{\circ}C$), acid(pH 2.0) and alkali(pH 11.0) treatment. The cytotoxic effect of isolated anticancer compounds on various cancer cell lines such as A549, SNU-1, KB, L1210, and Sarcoma 180 was investigated by MTT assay method. And these produced compounds also showed the broad antimicrobial spectrum to test strains such as bacteria and yeast.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.8
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• pp.1085-1089
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• 2006

The objectives of this study were to confirm a location of the calpastatin (CAST) gene in chromosome 2 and to detect associations of genetic variations with economic traits in the porcine CAST gene as a candidate gene for growth and meat quality traits in pigs. Calpastatin is a specific endogenous inhibitor of calpains. The calpain protease system is ubiquitous, and is involved in numerous growth and metabolic processes. Three single nucleotide variations were identified within a 1.6 kb fragment of the porcine CAST gene and these polymorphisms were used for genetic linkage mapping. Linkage and QTL mapping were performed with the National Livestock Research Institute (NLRI) reference families using eight microsatellites and SNP makers in the CAST gene. The porcine CAST gene was mapped adjacent to the markers, SW395 and SW1695 on SSC2 with LOD scores of 15.32 and 8.50, respectively. According to the QTL mapping, a significant association was detected at 82 cM between SW395 and CAST-Hinf I for weight at the age of 30 weeks. In addition, an association study was performed with the $F_2$ animals of NLRI reference families for Hinf I, Msp I and Rsa I polymorphisms in the CAST gene. Two polymorphisms, CAST-Rsa I and CAST-Hinf I, showed significant correlation for growth traits at p<0.01 and p<0.05, respectively.

• Journal of Microbiology and Biotechnology
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• v.20 no.10
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• pp.1403-1414
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• 2010

Aspergillus awamori BTMFW032, isolated from sea water, produced tannase as an extracellular enzyme under submerged culture conditions. Enzymes with a specific activity of 2,761.89 IU/mg protein, a final yield of 0.51%, and a purification fold of 6.32 were obtained after purification through to homogeneity, by ultrafiltration and gel filtration. SDS-PAGE analyses, under nonreducing and reducing conditions, yielded a single band of 230 kDa and 37.8 kDa, respectively, indicating the presence of six identical monomers. A pI of 4.4 and a carbohydrate content of 8.02% were observed in the enzyme. The optimal temperature was found to be $30^{\circ}C$, although the enzyme was active in the range of $5-80^{\circ}C$. Two pH optima, pH 2 and pH 8, were recorded, although the enzyme was instable at a pH of 8, but stable at a pH of 2.0 for 24 h. Methylgallate recorded maximal affinity, and $K_m$ and $V_{max}$ were recorded at $1.9{\times}10^{-3}$M and 830 ${\mu}Mol$/min, respectively. The impacts of a number of metal salts, solvents, surfactants, and other typical enzyme inhibitors on tannase activity were determined in order to establish the novel characteristics of the enzyme. The gene encoding tannase, isolated from A. awamori, was found to be 1.232 kb, and nucleic acid sequence analysis revealed an open reading frame consisting of 1,122 bp (374 amino acids) of one stretch in the -1 strand. In silico analyses of gene sequences, and a comparison with reported sequences of other species of Aspergillus, indicate that the acidophilic tannase from marine A. awamori differs from that of other reported species.

• BMB Reports
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• v.42 no.8
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• pp.529-534
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• 2009

Marbling score (MS) is the major trait that affects carcass quality in beef cattle. In this study, we investigated the association between genetic polymorphisms of the adipose differentiation-related protein gene (ADFP) and carcass traits in Korean cattle (also known as Hanwoo). Using direct DNA sequencing in 24 unrelated Korean cattle, 25 novel polymorphisms were identified within all exons and their flanking regions of ADFP, including the promoter region (1.5 kb). Among them, 21 polymorphic sites were selected for genotyping in the beef cattle (n = 425). Statistical analyses revealed that one promoter polymorphism (c.-56-18A > G) was associated with MS (P = 0.009). The 'A' allele of c.-56-18A > G exerted a lowering effect on MS, e.g., the lowest MS was found in 'A/A' (MS = 2.09 ${\pm}$ 1.23), intermediate in 'A/G' (MS = 2.11 ${\pm}$ 1.31), and the highest in 'G/G' (MS = 2.47 ${\pm}$ 1.47). Our findings suggest that these polymorphisms in ADFP might be important genetic factors involved in carcass quality in beef cattle.