• Title/Summary/Keyword: cyt b

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Toxin Genes and Antibiotic Resistance of Food Poisoning Bacteria Isolated from Food Service Equipment in Childcare Centers (어린이집 급식설비에서 분리된 식중독 미생물의 독소 유전자 및 항생제 내성)

  • Eun-yeong Kim;Chae-Young Kim;Ji-Yu Im;Jung-Beom Kim
    • Journal of Food Hygiene and Safety
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    • v.39 no.3
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    • pp.266-272
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    • 2024
  • This study assessed the contamination level of food poisoning bacteria on handles of food service equipment in childcare centers to prevent food poisoning and analyzed toxin genes and antibiotic resistance of isolated strains. The isolates used in this study were collected from 101 childcare centers in Jeollanam-do. Four strains of Bacillus cereus and two strains of Staphylococcus aureus were isolated on the handles of food service equipment (refrigerators and freezers). The toxin genes of B. cereus were detected as nheA, nheB, nheC, entFM, and cytK. No toxin genes of S. aureus were detected. B. cereus showed resistance to β-lactam antibiotics, such as ampicillin and cefepime. S. aureus also showed resistance to antibiotics such as ampicillin and cefepime. Therefore, microbial safety and hygiene management, such as periodic sterilization of handles, should be strengthened to prevent food poisoning caused by cross-contamination of food service equipment handles in childcare centers.

Expression of Mosquitocidal Bacillus sphaericus Binary Toxin and B. thuringiensis cry11B Genes in B. thuringiensis 407

  • Park, Hyun-Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.2
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    • pp.185-189
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    • 2001
  • Wild type Bacilus thuringiensis subsp. israelensis and B. sphaericus toxins have been used separately as active in ingredients for bacterial insecticides to control mosquito larvae due to their comparable toxicity to chemical insecticides. Cry11B, recently cloned from B. thuringiensis subsp. jegathesan, shows higher toxicity against three major species of mosquito larvae than Cry11A, one of the major component of B. thuringiensis subsp. israelensis inclusion body. To determine whether the combination of cry11B and B. sphaericus binary toxins is as toxic as B. thuringiensis subsp. israelensis parental strain, cry11B and B. sphaericus binary toxins genes were co-expressed as an operon using cytlA promoters/STAB-SD hybrid expression system in B. thuringiensis subsp. israelensis acrystalliferous strain 4Q7. However, unexpectedly, B. sphaericus binary toxins were barely produced, whereas relatively large amount of Cry11B was produced. When this strain was grown in four different media, NB+G and Peptonized Milk produced more toxin proteins and spores per unit of media than GYS and G-Tris. Toxicity of this strain against fourth instar Culex quinquefasciatus was ranged from of 8.3 to 45.7 ng/ml, with NB+G culture being the highest, and GYS culture was the lowest.

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Study on the Genetic Variation of the Mitochondrial DNA in the Beet Armyworm, Spodoptera exigua (H bner), Using PCR-RFLP (PCR-RFLP를 이용한 파방나방 (Spodoptera exigua(H bner)) 미토콘트리아 DNA의 유전변이 연구)

  • 김용균;이명렬;정충렬
    • Korean journal of applied entomology
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    • v.37 no.1
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    • pp.23-30
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    • 1998
  • Restriction fragment length polymorphism (RFLP) of a DNA has been a useful tool for analyzing genetic variation. This research was performed to establish an RFLP analytic method on the mitochondrial DNA (mtDNA) of the beet armyworm, Spodoptera exigua (Hiibner). To do this, total size of the mtDNA was measured and polymerase chain reaction (PCR) primers were selected. Its mitochondrial genome size was ca. 16kb. From a serial PCR test of 29 primers refered to the compilation of Simon et al. (1994), 22 primers were selected to amplify its mtDNA fragments. These primers resulted in short (300-700 bp) or long (1000-2000 bp) DNA products which represented a total or partial sequence of each of CO-I, CO-11, Cyt-B, ND-1, 12s rRNA, 16s rRNA, and some tRNAs. PCR-RFLP was performed in some variable mtDNA regions with 8 kinds of 4bp recognizing restriction enzymes. Different populations from Andong, Kyungsan, and Sunchun did not show any restriction site polymorphisms but had some length variation in certain regions of mtDNA.

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Introduction of Stay Green Mutant for the Development of Black Seed Coat and Green Cotyledon Soybean Variety (녹색자엽 검정콩 품종 육성을 위한 Stay green 변이체 활용)

  • Kang, Sung-Taeg;Seo, Min-Jung;Moon, Jung-Kyeong;Yun, Hong-Tae;Lee, Young-Ho;Kim, Si-Ju;Hwang, Young-Sun;Lee, Suk-Ki;Choung, Myoung-Gun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.55 no.3
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    • pp.187-194
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    • 2010
  • The soybean stay green mutant genotype (SSG) derived from the nuclear gene, d1d2, and cytoplasmic gene, cytG, inhibit the breakdown of chloroplast in the leaves, pod walls, seed coats, and embryos during maturity. Soybean seed with black seed coat and green cotyledon (SBG) are preferred than black seed coat with yellow cotyledon (SBY) especially for cooking with rice and as source of traditional food in Korea. The researchers evaluated the seed's chlorophyll content of SSG and introduced SSG to the SBG variety breeding program. The seed chlorophyll content of SSG with d1d2 was $39.93{\sim}60.80\;{\mu}g/g$ and SSG with cytG $38.08{\sim}39.89\;{\mu}g/g$. The Korean SBG variety which was derived from SSG with cytG, contains $16.35{\sim}37.73\;{\mu}g/g$. The composition of seed chlorophyll differs according to the genetic background of SSG genotype. Inheritance study showed that cotyledon color was segregated 15:1 (yellow:green) at $F_2$ seed indicating two recessive genes control green cotyledon as revealed by previous study. Only less than 3% soybean lines showed black seed coat with green cotyledon among crosses SBY and SSG (d1d2). Results showed that SSG with d1d2 can be used as a good source for SBG with high chlorophyll content in the seed cotyledon, but due to the complex genetic behavior, breeding resource of SBG with d1d2 should be prepared to improve the breeding efficiency for development SBG variety.

Discordance between Morphological and Molecular Variations of the Genus Macroramphosus (Macroramphosidae) from Korea (한국산 대주둥치속(대주둥치과) 어류의 형태와 분자 변이의 불일치)

  • Sohn, Min-Soo;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.32 no.4
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    • pp.199-209
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    • 2020
  • In order to clarify the taxonomic status of the Korean Macroramphosus species, which were previously confused, we investigated morphological and molecular variations of Macroramphosus (18 individuals) from Korea, and Macroramphosus (35 individuals) from Japan and Taiwan, and compared with those of M. scolopax from type locality (Mediterranean Sea). Although the Korean and Japanese specimens of Macroramphosus were clearly divided into two types in the first dorsal spine length (22.8~32.1% in A-type vs. 15.6~21.4% in B-type), distance between the first dorsal fin and second dorsal fin (6.4~9.7% vs. 8.6~13.3%), and body depth (20.0~28.0% vs. 17.3~22.6%), no genetic differences among all individuals of longspine snipefish between them were found at the specific level [d=0.0~3.3% in control region (CR); 0.0~1.3% in cytochrome b (cytb); 0.0~0.5% in cytochrome c oxidase subunit I (COI)]. Whereas, they were well distinguished in genetics (9.9~11.5% in CR; 3.8~4.6% in cytb; 1.2~3.6% in COI) from those of M. scolopax in Mediterranean Sea. It needs the scientific name of the longspine snipefish (M. scolopax) in Korea be changed as M. japonicus (and/or M. sagifue). However, our results could not find evidence of consistency between morphological and mitochondrial DNA variations which suggests that their differentiation event may occur fairly recently. Further studies using more sensitive markers such as microsatellite are needed to clarify the degree of gene flow between the two types.

Application for Identification of Food Raw Materials by PCR using Universal Primer (일반 프라이머를 이용한 PCR의 식품원료 진위 판별에 적용)

  • Park, Yong-Chjun;Jin, Sang-Ook;Lim, Ji-Young;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Han, Sang-Bae;Lee, Sang-Jae;Lee, Kwang-Ho;Yoon, Hae-Seong
    • Journal of Food Hygiene and Safety
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    • v.27 no.3
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    • pp.317-324
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    • 2012
  • In order to determine an authenticity of food ingredient, we used DNA barcode method by universal primers. For identification of animal food ingredients, LCO1490/HCO2198 and VF2/FISH R2 designed for amplifying cytochrome c oxidase subunit1 (CO1) region and L14724/H15915 for cytochrome b (cyt b) region on mitochondrial DNA were used. Livestock (cow, pig, goat, sheep, a horse and deer) was amplified by LCO1490/HCO 2198, VF2/FISH R2 and L14724/H15915 primers. Poultry (chicken, duck, turkey and ostrich) was amplified by LCO1490/HCO 2198 and VF2/FISH R2 primers. But, Fishes (walleye pollack, herring, codfish, blue codfish, trout, tuna and rockfish) were only amplified by VF2/FISH R2 primers. For plant food ingredients, 3 types of primers (trnH/psbA, rpoB 1F/4R and rbcL 1F/724R) have been used an intergenic spacer, a RNA polymerase beta subunit and a ribulose bisphosphate carboxylase region on plastid, respectively. Garlic, onion, radish, green tea and spinach were amplified by trnH/psbA, rpoB 1F/4R and rbcL 1F/724R. The PCR product sizes were same by rpoB 1F/4R and rbcL 1F/724R but, the PCR product size using trnH/psbA primer was different with others for plants each. We established PCR condition and universal primer selection for 17 item's raw materials for foods and determine base sequences aim to PCR products in this study. This study can apply to determine an authenticity of foods through making an comparison between databases and base sequences in gene bank. Therefore, DNA barcode method using universal primers can be a useful for species identification techniques not only raw materials but also processed foods that are difficult to analyze by chemical analysis.

Molecular Identification and Distribution Pattern of Fish Eggs Collected around Jejudo Island (제주 연안해역 어란의 분자동정과 분포양상)

  • Han, Song-Hun;Kim, Maeng Jin;Song, Choon Bok
    • Korean Journal of Ichthyology
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    • v.27 no.4
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    • pp.284-292
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    • 2015
  • Monthly variation of species composition and abundance of fish eggs were examined to know the spawning time and location of the fishes inhabiting the coastal region of Jejudo Island. Samplings had been performed at the four locations (Jeju port, Seongsanpo, Seogwipo port and Chagwido) with a bongo net which was towed monthly at the sea surface from August 2006 to July 2007. The fish eggs were identified based on phylogenetic analyses with the DNA sequences generated through PCR-amplification and sequencing of the mitochondrial cytochrome b gene. Among a total of 43 taxa classified during the study period, 34 were identified to species, 4 to families, and the remaining 5 were unidentified. Of them, 23 taxa were occurred at Jeju port, 21 at Seongsanpo, 19 at Seogwipo port and 18 at Chagwido, whereas 15 taxa were found in September 2006, 12 in June 2007, 6 to 8 in August 2006 and January~May 2007, 5 in each October and November 2006, 3 in each December 2006 and July 2007. Among 34 species of fish eggs, Engraulis japonicus and Callanthias japonicus most frequently appeared at 16 times out of 48 observations in total. When those eggs were collected, the surface seawater temperature ranged $14.0{\sim}28.6^{\circ}C$ for E. japonicus and $14.9{\sim}20.5^{\circ}C$ for C. japonicus. The success rates of PCR-amplification and species identification in this study were 68.3% and 79.1%, respectively.

The distribution of $^{14}C$-warfarin and the purification of hepatic microsome induced isozymes with coumarin ($^{14}C$-warfarin의 분포 및 쿠마린 유도체류에 의하여 간에서 유도된 동위효소의 정제)

  • Park, Sung-Woo;Kim, Eun-Ho;Min, Ji-Sook;You, Jae-Hoon;Lee, Hee-Sung;Seo, Bae-Seck;Han, Wan-Soo
    • Analytical Science and Technology
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    • v.5 no.1
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    • pp.83-90
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    • 1992
  • The $^{14}C$-warfarin used as rodenticids was identified from various organs of sprague dawley with scintillation counter. And the cytochrome p-450 which was induced by coumarin derivatives was identified with electrophoresis. The distributions of $^{14}C$-warfarin after $14.8{\mu}Ci/kg$ oral application at each organ was as follows; urine-7.5%, blood-0.44%, feces-0.9%, liver-0.66%, lung-0.86%, kidney-0.8%, heart-0.43% and spreen-0.33% after 24hrs. The cytochrome p-450 was purified by Octyl Sepharose CL-4B hydrophobic chromatography and isozymes were 50.8 Kd in control group, 53.3 Kd and 55.2 Kd in coumarin pretreated group and 50.8 Kd, 54.6 Kd and 57.7 Kd in warfarin pretreated group.

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STRUCTURAL PERTURBATIONS INDUCED BY PHOTODYNAMIC ACTION OF PORPHYRIN AGGREGATES ON PLASMA MEMBRANE AND MICROSOMES OF GLIOBLASTOMA CELLS

  • Sreentvasan, Rajesh;Joshi, Preeti G.;Joshi, Nanda B.
    • Journal of Photoscience
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    • v.4 no.2
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    • pp.41-48
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    • 1997
  • The plasma membrane and microsomes, isolated from the cells treated with hematoporphyrm derivative (HpD) for 1 and 24 h, accumulated the aggregated porphyrin. The quantity of aggregated porphyrin was same in the plasma membrane and microsomes after isolating them from cells treated with HpD for 1 h whereas the microsomes accumulated higher quantity of aggregated porphyrin when cells were treated with HpD for 24 h. Photodynamic action of aggregated porphyrin on plasma membrane and microsomes was investigated using lipid specific fluorescent probes: 1,6-diphenyl-1,3,5-hexatrine (DPH) and 1-(4-trimethylammonium), 6-diphenyl-1,3,5-hexatrine(TMA-DPH). The time dependent anisotropy of these probes in the membranes was measured and the decay of anisotropy was analyzed using wobbling in cone model. Upon irradiation both the plasma membrane and the microsomes showed an increase in the limiting anis~)tropy and order parameter and a decrease in the cone angle of the lipid probes. The increase in the limiting anisotropy was pronounced in membranes isolated from the cells treated with HpD for 24 h. Photoinduced change in the limiting anisotropy was dependent on the duration of incubation of cells with HpD before isolating the membranes. In both the membranes. the membrane core was affected more as compared to the outer leaflet. In addition to the structural changes, a decrease in Na$^+$-K$^+$-ATPase and NADPH cyt c reductase activity was also observed upon irradiation of HpD treated cells. Inhibition in NADPH cyt c reductase was more when cells were treated with HpD for 24 h, however, Na$^+$-K$^+$-ATPase activity did not depend on the duration of the treatment of cells with HpD before irradiation. Our results suggest that the extent of photoinduced perturbations in the membranes varies as a function of duration of the treatment of cells with HpD and the membrane core is more susceptible to the photodynamic action of aggregated porphyrin.

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Structural Analysis of Cu Binding Site in [Cu(I)·d(CpG)·d(CpG)-2H]-1 Complex

  • Im, Yu-Jin;Jung, Sang-Mi;Kang, Ye-Song;Kim, Ho-Tae
    • Bulletin of the Korean Chemical Society
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    • v.34 no.4
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    • pp.1232-1236
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    • 2013
  • The Cu cation binding sites of $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex have been investigated to explain the $[Cu{\cdot}DNA]$ biological activity caused by the Cu association to DNA. The structure of $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex was investigated by electrospray ionization mass spectrometry (ESI-MS). The fragmentation patterns of $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex were analyzed by MS/MS spectra. In the MS/MS spectra of $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex, three fragment ions were observed with the loss of d(CpG), {d(CpG) + Cyt}, and {d(CpG) + Cyt + dR}. The Cu cation binds to d(CpG) mainly by substituting the $H^+$ of phosphate group. Simultaneously, the Cu cation prefers to bind to a guanine base rather than a cytosine base. Five possible geometries were considered in the attempt to optimize the $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex structure. The ab initio calculations were performed at B3LYP/6-31G(d) level.