• Journal of Soil and Groundwater Environment
• /
• v.19 no.6
• /
• pp.59-71
• /
• 2014

We investigated soil contamination depending on the land use by examining the contamination levels and distribution characteristics of 16 polycyclic aromatic hydrocarbons (PAHs) in the national soil. Total PAHs (the sum of 16 PAH concentrations) and carcinogenic PAHs (the sum of seven carcinogenic PAH concentrations) were $8.50{\sim}3,437.16{\mu}g/kg$ and $2.94{\sim}2,136.96{\mu}g/kg$, respectively. The concentration of benzo(a)pyrene, one of the contaminants regulated by the soil quality standard in the nation, was $ND{\sim}924.73{\mu}g/kg$. Its maximum value of $924.73{\mu}g/kg$ was detected in railroad (Region 3) and is approximately 13% of the standard value for Region 3 (i.e., 7 mg/kg). We also investigated the characteristics of contamination sources of PAHs in soil of the upland, forests, roads, and railroads, examining the fraction distribution of PAHs concentration by the number of benzene rings against the total PAHs concentration. The results demonstrate that the mean fraction of 4~6-ring PAHs against total PAHs concentration in soil was in the range of 51.8~80.7% with relative abundance of high-molecular PAHs, showing that the origin of contamination is under the category of combustion sources. When the molecular indices (Flu/(Flu/Pyr), Ant/(Ant+Phe), InP/(InP+BP), and BaA/(BaA+Chr)) were applied, they were also categorized as petroleum-based combustion sources. The individual PAH concentrations in soil by the land use were grouped into Regions 1, 2, and 3, which are statistically treated and are the parts of the national category system of soil quality standard. As a result, the concentration level of 16 PAHs was $0.02{\sim}2.63{\mu}g/kg$ in Region 1, $0.05{\sim}4.26{\mu}g/kg$ in Region 2, and $2.36{\sim}178.27{\mu}g/kg$ in Region 3. The concentration level of 6 carcinogenic PAHs was 14.2~320.4% against that of benzo(a)pyrene in Region 3 and sites of recycling waste sleepers. Considering that there were similarities among PAHs in terms of structures and toxicities, it would be recommended to review other types of carcinogenic PAHs together with benzo(a)pyrene when developing the soil quality standards in the nation.

• Applied Biological Chemistry
• /
• v.50 no.2
• /
• pp.95-100
• /
• 2007

The cellulase gene of Bacillus licheniformis K11 which has plant growth-promoting activity by auxin and antagonistic ability by siderophore was cloned in pUC18 using PCR employing heterologous primers. The 1.6kb PCR fragment contained the full sequence of the cellulase gene, denoted celW which has been reported to encode a 499 amino acid protein. Similarity search in protein data base revealed that the cellulase from B. licheniformis K11 was more than 97% identical in amino acid sequence to those of various Bacillus spp. The cellulase protein from B. licheniformis K11, overproduced in E. coli DH5${\alpha}$ by the lac promoter on the vector, had apparent molecular weight of 55 kDa upon CMC-SDS-PAGE analysis. The protein not only had enzymatic activity toward carboxymethyl-cellulose (CMC), but also was able to degrade insoluble cellulose, such as Avicel and filter paper (Whatman$^{\circledR}$ No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. Consequently B. licheniformis K11 was able to suppress the peperblight causing P. capsici by its cellulase. Biochemical analysis showed that the enzyme had a maximum activity at 60$^{\circ}C$ and pH 6.0. Also, the enzyme activity was activated by Co$^{2+}$ of Mn$^{2+}$ but inhibited by Fe$^{3+}$ or Hg$^{2+}$. Moreover, enzyme activity was not inhibited by SDS or sodium azide.

• Korean Journal of Microbiology
• /
• v.42 no.1
• /
• pp.40-46
• /
• 2006

Galactose joined to glucose by a $\beta(1\rightarrow4)$ glycosidic bond makes lactose and this disaccharide is rich in milk. It is known that lacotse is hydrolyzed to each monomeric sugar by either lactase in human or $\beta-galactosidase$ in bacteria. Ingestion of milk by lactase-deficient persons causes a temporary diarrhea and subsequent chronic diarrhea results in colitis with chronic inflammation. We isolated a $\beta-galactosidase$ producing psycrotolerant strain AS-20 from near cattle shed and investigated the growth at various temperature conditions. Whereas Escherichia coli strains did not grow at $10^{\circ}C$, the AS-20 strain could grow well at this low temperature and showed optimal growth at $30^{\circ}C$. The isolated strain was identified as 97% Hafnia alvei by biochemical properties. This strain could ferment glucose, lacotse, maltose, mannitol, xylose, ONPG, rhamanose and L-arabinose, and decarboxylate lysin and ornithine. To confirm the identity of isolated strain we amplified 16S rDNA by PCR and searched similarity of the 1426 bp DNA sequcence with Genbank database. The strain AS-20 showed 99% similarity with Hafnia alvei. The activity of $\beta-galactosidase$ was 1.5 times higher when the cell was grown at 10 or $20^{\circ}C$ than at $30^{\circ}C$. The highest enzyme activity of AS-20 was also much higher than that of E. coli, which was grown at $30^{\circ}C$.

• Journal of Animal Science and Technology
• /
• v.44 no.6
• /
• pp.653-660
• /
• 2002

We considered KIT gene as a candidate gene for the white-spotting pattern in cattle. This study was carried out to detect genetic variation of c-KIT receptor gene and to investigate association between the mutation and the white-spotting pattern in cattle. PCR-RFLP analysis within intron 6 of c-KIT receptor gene were performed with 8 cattle breeds including Hanwoo, Angus, Brown Swiss, Charolais, Hereford, Holstein, Limousin and Simmental. When PCR product of approximately 2,440 bp including intron 6 of c-KIT receptor gene was sequenced, four nucleotide substitutions were found within intron 6 of the bovine c-KIT receptor gene. In PCR-RFLP analysis, three alleles (A, B and C), two alleles (A and B) and two alleles (A and B) at each locus were identified by MspⅠ, BsrBⅠ and NdeⅠ, respectively. Although frequencies of allele at each locus were different among cattle breeds, we could not get any evidence related with white or white spotting phenotypes in these mutations on intron 6 of c-KIT receptor gene. However, we can not entirely exclude the possibility that c-KIT receptor gene is responsible for white spotting phenotype in cattle. Thus, further studies need to detect other mutations in c-KIT receptor gene and to test association of those mutations and coat color phenotypes in cattle.

• Journal of Life Science
• /
• v.24 no.4
• /
• pp.343-351
• /
• 2014

Phosphoinositide 3-kinase (PI3K) plays a central role in cell signaling and leads to cell proliferation, survival, motility, exocytosis, and cytoskeletal rearrangements, as well as specialized cell responses, superoxide production, and cardiac myocyte growth. PI3K is divided into three classes; type I PI3K is preferentially expressed in leukocytes and activated by ${\beta}{\gamma}$ subunits of heterotrimeric G-proteins. In this study, the cDNAs encoding the $PI3K{\gamma}$ gene were isolated from a brain cDNA library constructed using the flounder (Paralichthys olivaceus). The sequence of the isolated $PI3K{\gamma}$ was 1341 bp, encoding 447 amino acids. The nucleotide sequence of the $PI3K{\gamma}$ gene was analyzed with that of other species, including Oreochromis niloticus and Danio rerio, and it turned out to be well conserved during evolution. The $PI3K{\gamma}$ gene was subcloned into the expression vector pET-44a(+), and expressed in the E. coli BL21 (DE3) codon plus cell. The resulting protein was expressed as a fusion protein of approximately 49 kDa containing a C-terminal six-histidine extension that was derived from the expression vector. The expressed protein was purified to homogeneity by His-tag affinity chromatography and showed enzymatic activity corresponding to $PI3K{\gamma}$. The binding of wortmannin to $PI3K{\gamma}$, as detected by anti-wortmannin antisera, closely followed the inhibition of the kinase activities. The results obtained from this study will provide a wider base of knowledge on the primary structure and characterization of the $PI3K{\gamma}$ at the molecular level.

• Journal of Life Science
• /
• v.31 no.3
• /
• pp.356-365
• /
• 2021

Recently, we sequenced the entire genome of a freshwater agar-degrading bacterium Cellvibrio sp. KY-GH-1 (KCTC13629BP) to explore genetic information encoding agarases that hydrolyze agarose into monomers 3,6-anhydro-L-galactose (L-AHG) and D-galactose. The KY-GH-1 strain appeared to possess nine β-agarase genes and two α-neoagarobiose hydrolase (α-NABH) genes in a 77-kb agarase gene cluster. Based on these genetic information, the KY-GH-1 strain-caused agarose degradation into L-AHG and D-galactose was predicted to be initiated by both endolytic GH16 and GH86 β-agarases to generate NAOS (NA4/NA6/NA8), and further processed by exolytic GH50 β-agarases to generate NA2, and then terminated by GH117 α-NABHs which degrade NA2 into L-AHG and D-galactose. More recently, by employing E. coli expression system with pET-30a vector we obtained three recombinant His-tagged GH50 family β-agarases (GH50A, GH50B, and GH50C) derived from Cellvibrio sp. KY-GH-1 to compare their enzymatic properties. GH50A β-agarase turned out to have the highest exolytic β-agarase activity among the three GH50 isozymes, catalyzing efficient NA2 production from the substrate (agarose, NAOS or AOS). Additionally, we determined that GH117A α-NABH, but not GH117B α-NABH, could potently degrade NA2 into L-AHG and D-galactose. Sequentially, we examined the enzymatic characteristics of GH50A β-agarase and GH117A α-NABH, and assessed their efficiency for NA2 production from agarose and for production of L-AHG and D-galactose from NA2, respectively. In this review, we describe the benefits of recombinant GH50A β-agarase and GH117A α-NABH originated from Cellvibrio sp. KY-GH-1, which may be useful for the enzymatic hydrolysis of agarose for mass production of L-AHG and D-galactose.

• Korean journal of applied entomology
• /
• v.47 no.4
• /
• pp.333-336
• /
• 2008

During the excavation to find ancient rice seeds from the peat layer (12,500-14,800 bp based on 14 dating samples) of Soro-ri, Cheongwon-gun near Cheongju city, we found not only old rice seeds, but also other organic materials including insect fossil skeletons. Although no complete specimen was found in this study, some body parts were diagnotic enough for identifying species. Among the identified insect fossils, two species were successfully identified based on their characteristic forewing morphologies, e.g., cross-ridges on forewing. The two identified species, Plateumaris sericea and Donacia flemola or near, belong to Donaciinae in Chrysomelidae, and their larvae are known to feed on the root of wetland plants, mainly in Poaceae or Cyperaceae. Such finding proved that a wetland environment was maintained in Soro-ri at that time, and was suitable for such insects to survive. Therefore, evidence from the fossilized rice seeds, together with these insect fossils, clearly proved that the environment at that time was suitable for rice, a common Poaceae species, growing.

• Proceedings of the KSAR Conference
• /
• 2001.03a
• /
• pp.58-58
• /
• 2001

본 연구는 섬유소 분해효소 유전자 Cel D(Cellulase Digestion)가 도입된 형질전환 돼지 생산을 통하여 섬유소 함유 사료의 이용 효율을 증대시키고, 나아가 장기이식동물 및 고가의 의료용 단백질 생산가축을 개발하기 위한 원천기술 확보에 있다. 섬유소분해 유전자의 크로닝 및 조직 특이적 발현벡터를 개발하기 위하여, 우선 소의 위중 제4위 내의 미생물로부터 전체 유전자를 분리하였고, 이렇게 작성된 DNA library에서 섬유소분해 관련 유전자인 약 2.0 kb의 Cel D유전자를 크로닝하였으며, 췌장 특이적 발현 프로모터(rat elastase I: 약 200bp)를 크로닝한 후, 미세주입용 형질전환 재조합 벡터를 구축하기 위하여 rElastase I 프로모터 하류에 섬유소 분해 유전자(Cel D)를 연결하여 약 3.0 kb 크기의 재조합 벡터를 준비하였으며, 재조합 유전자를 1세포기 수정란 전핵내에 미세주입 하기 위해 Sal I과 BglII를 이용 유전자 단편을 만들었다. 구축된 유전자를 미세주입하기 위한 수정란을 회수하기위해 총68두의 돼지를 4-5두씩 분리사육하면서 발정동기화 및 과배란 유기를 위해 PG600, Altrenogest, FSH, hCG를 투여하였으며 hCG투여후 약54시간에 외과적 방법에 의해 총 1,359개의 수정란을 회수하였고, 이중 미세주입가능한 1세포기 수정란은 1,296개로 두당 평균 15.9개 였다. 1,296개의 1세포기 수정란 중에서 재조합 유전자(rE I-CelD)가 미세주입된 660개의 수정란을 32두의 수란돈에 외과적 방법에 의해 이식하였으며, 이식되어진 모돈 13두가 분만하여 40.6%의 임신율을 나타내었다. 이렇게 분만된 13두에서 총 65두(암:33두, 수:32두)의 자돈이 생산되었으며, 형질전환 여부를 판명하기 위해 자돈의 꼬리조직으로 부터 genomic DNA를 추출하고 PCR 검정을 실시하였다. PCR 검정 결과, 섬유소 분해 유전자가 도입된 자돈은 5두 이었으며, 그 결과를 Table에 나타내었다.(Table Omitted) Table 1 에서와 같이 섬유소 분해효소유전자가 형질전환된 자돈은 65마리 중 5마리로 7.69%의 형질전환율을 나타내었으며, 5마리의 자돈중 2두(암:1두, 수:1두)는 분만 후 즉시 폐사되었으며 2두(암:1두, 수:1두)는 86일령 그리고 14일령에 폐사하여 현재 1두(암)가 생존하여 섬유소 분해 사양 실험 중에 있다.

• KSBB Journal
• /
• v.17 no.4
• /
• pp.370-375
• /
• 2002

Isolation of a gene and determination of its expression pattern are essential in understanding its function. Among the genes localized in 12ql3, stSG3435 EST was chosen to study its expression pattern. The full-length CDNA was cloned by screening of human brain CDNA library and its sequence was determined by serial deletion followed by automated sequencing of the clones with overlapping fragments. The sequence analysis revealed that stSG 3435 CDNA displayed 100% identity to human MYGI and 86% identity to mouse melanocyte proliferation gene-1 (Gamm 1) originally identified from melanocyte, suggesting that MYGI determined by Northern blot analysis revealed the strongest expression in testes with ubiquitous expression in all the tissues tested. In order to investigate the cellular localization of its protein product, the green fluorescence protein gene was fused into the full-length coding sequence of MYGI, Transfection of the fusion construct followed by confocal microscopy resulted in the green fluorescence signal as a punctate state in cytoplasm indication that MYGI was localized in one of the cellular organelles.

• Korean Journal of Veterinary Service
• /
• v.31 no.1
• /
• pp.1-16
• /
• 2008

Salmonella infections cause the disease in pigs but also some zoonotic Salmonella serotypes can be transmitted to human through swine products, resulting in food poisoning. The objective of this study was to investigate the bacteriological prevalence and detection of invA gene using Salmonella specific polymerase chain reaction (PCR), the epidemiological characteristics related to Salmonella strains cultured from pig samples in Gangwon areas using serotyping, random amplified polymorphic DNA (RAPD) and pulsed field gel electrophoresis (PFGE) methods. During the period of November 2001 through April 2002, 1,174 ileocecal lymph node were collected from the slaughtered pigs raised in 38 farms located in Gangwon province. The samples were submerged in boiling water and macerated in saline and lymph node homogenates were inoculated into Tetrathionate broth with iodine (TTB, Difco, 0.5% iodine was added) for enrichment growth. Then additional tests were performed using several mediums, and suspects were identified by API 20E kit (BioMerieux) and PCR. Of total 1,174 samples from 38 farms, 44 (3.7%) were isolated as Salmonella spp from 13 farms (34.2%). Of 44 isolates, 31 were in Yangyang region, followed by 9 in Goseong, 2 in both Gangneung and Sokcho. However, there was no difference in regional isolation frequency. All isolates have a 521bp amplified product in Salmonella specific PCR with primer invA which encodes in proteins for invasion of epithelial cells. Of 44 recovered serotypes, 23 (52.3%) were S Eingedi, 10 (22.7%) S Schwarzengrund, 9 (20.5%) S Typhimurium, and 2 (4.5%) S Mbandaka. In RAPD analysis, there appeared to be unique bands distinguishing each serotype, although similarities exist between the different serotypes. Four serotypes of 44 Salmonella isolates appeared to fall into 14 different RAPD types. In PFGE analysis, 9 S Typhimurium were tested with XbaI enzyme and SpeI enzyme. The combination of results obtained with two enzymes subdivided the 9 S Typhimurium into 4 PFGE types.