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Protein Patterns of Ovary and Hemolvmph in Gerris paludum

  • 황명실;최원철
    • The Korean Journal of Zoology
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    • v.32 no.4
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    • pp.337-350
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    • 1989
  • In this study, we compared patterns of hemolvmph with ovarian proteins during the yolk formation of Gewis paludum by using SDS/polyacrylamide gel electrophoresis and he dimen-tlonal electrophoresis. In addition, we examined patterns of glycoprotein which were composed of yolk substances. The results were as follows: The protein patterns in ovary of the 5th instar without eggs were similar to those of adult after ovulation. Protein amounts of the ovaw without developing eggs were less than those of the ovaw containing oocvtes or matured eggs at the molecular weights range from 66, 000 to 110.000 daltons. No glvcoproteins were observed in ovaw without eggs. But the glvcoproteins were gradually increased according to development of eggs in the 5th instar. After the ovulated ovaw of adult, no glvcoprotein bands urere occurred as the bands of the ovary without eggs in the 5th instar. Also, amounts of hemolvmph protein between 66, 000 and 110, 000 in molecular weight were increased during yolk formation of the 5th instal. The results suggest that ovarian protein substances may originate from hemolpnph. In the 5th instar, the amounts of glycoprotein of developing eggs was gradually increased in the hemolymph. The band of M.W. 29, 500 was occurred in the hemolpnph of the 5th instar and adult without eggs. This protein mal be precursor of glvcoprotein in the high molecular weight area in the ovary of more developed eggs. The number of spots and the amounts of protein in ovary without eggs were less than those of ovary containing eggs by two dimensional elec-trophoresis. The protein bands between 45, 000 and 110, 000 almost appeared in acidic field of he dimensional gel. Especially, the band, M.W. 109, 000, uras separated 3 spots, a1, a2, and as. The band, M.W. 102, 000, has spots which designated b1, b2, b3, and b4. Besides, proteins below M.W. 24, 000 were occurred less spots in the basic field than those of acidic field. The mechanism of intracellular organs (= cell organells) uras related to the yolk protein synthesis of oocyte in the process of yolk protein formation of derris pcfudum.

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The Pulation Structure of the Pacific Cod (Gadus macrocephalus Tilesius) Based on Mitochondrial DNA Sequences (미토콘드리아 유전자 염기서열 분석에 의한 대구 계군 분석)

  • Seo, Young-Il;Kim, Joo-Il;Oh, Taeg-Yun;Lee, Sun-Kil;Park, Jong-Hwa;Kim, Hee-Yong;Cho, Eun-Seob
    • Journal of Life Science
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    • v.20 no.3
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    • pp.336-344
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    • 2010
  • To assess population structure and genetic diversity among the Pacific cod (Gadus macrocephalus), we investigated mtDNA COI gene sequences of 7 populations. Samples were obtained from Sokcho, Wolsung, Geojedo, Yeosu, Geomundo and Westsouth in 2008 and 2009 (n=28). The sequence analysis of 28 individual samples showed 8 haplotypes, ranging in sequence divergence by pairwise comparisons from 0.2 to 2.2% (1 bp-11 bp). The Gal haplotype was found in Wolsung, Geojedo, Yeosu, Geomundo and Westsouth, and was regarded as the main haplotype of Korean Pacific cod. Ga2, Ga3, Ga6 and Ga7 haplotypes were found only in Sokcho. In the PHYLIP analysis, 8 haplotypes formed two independent groups: cladeA consisted of Ga2, Ga3, Ga6 and Ga7 haplotypes, whereas cladeB contained Gal, Ga4, Ga5 and Ga8 haplotypes. The genetic relationship between the two groups was weakly supported by bootstrap analysis(<50%). In pairwise comparisons between 6 populations other than that from Sokcho, a very high per generation migration ratio ($N_m$=infinite) and a very low level of geographic distance ($F_{sr}=-0.0123-(-0.0423)$) were observed. The estimates of genetic distance between Sokcho and the other localities were all statistically significant (p<0.05, p<0.01, p<0.001), indicating a limited mtDNA-based gene flow between Sokcho and other regions. The finding of the lowest genetic diversity in the Sokcho population (nucleotide diversity=0.00589) may be a result of relatively small population size and interrupted gene flow to other localities. Consequently, the overall considerable migration of Pacific cod population in Korea caused a genetically homogeneous structure to form, although a distinct population was found in this study.

Physiological and Biochemical Characterization of Bacillus spp. from Polychaete, Perinereis aibuhitensis (갯지렁이(Perinereis aibuhitensis)에서 분리한 Bacillus spp.의 생리생화학적 특성 분석)

  • Shin, Seyeon;Yundendorj, Khorloo;Lee, Sang-Suk;Kang, Kyoung-Ho;Kahng, Hyung-Yeel
    • Journal of Life Science
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    • v.23 no.3
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    • pp.415-425
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    • 2013
  • This study compared the characteristics of five Bacillus strains capable of aerobic and anaerobic growth, CBW3, CBW4, CBW9, CBW14 and EBW10. They were isolated and selected from a polychaete, Perinereis aibuhitensis, which is known as a good degrader of organic compounds in marine wetland. Based on a 16S rRNA sequence, CBW3 and CBW14 were found to share more than 99.8% similarity with B. nanhaiensis, B. arsenicus and B. barbaricus. CBW4, CBW9 and EBW10 shared 92.7%, 99.8%, and 99.8% similarity with B. anthracis, B. algicoa and B. thuringiensis, respectively. The temperature, salinity, and pH ranges of the cell growth of the Bacillus strains were $4-45^{\circ}C$, 0-17%, and pH 5-pH 9, respectively. All Bacillus strains were found to exhibit enzyme activities for the degradation of casein and starch. Notably, strain EBW10 exhibited the enzyme activities for all the tested macromolecules, DNA, casein, starch, cellulose, and four kinds of Tweens, which suggests the possibility that it had protease, amylase, cellulose, and lipase. All five Bacillus strains had alkaline phosphatase activities, and the strains CBW3, CBW4, and EBW10 also had acid phospatase. Strains CBW3 and EBW10 exhibited the enzyme activities both for esterase (C4) and esterase lipase (C8). The analysis of fatty acids revealed that in all strains, major fatty acids were anteiso $C_{15:0}$ and iso $C_{15:0}$.

Effects of Medicinal Plants Extract on Naengmyeon Broth (약용식물 추출물이 냉면육수에 미치는 영향)

  • 김명숙;최윤희;홍선표
    • The Korean Journal of Food And Nutrition
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    • v.16 no.4
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    • pp.328-333
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    • 2003
  • When the extract of the medicinal plants, Kaempferia galanga L., Zanthoxylum bungeanum, Eugenia caryophyllata, Foeniculum vulgare, was added to Naengmyeon broth with the concentration of 0.1% and 0.3% each, its effect during the preservation time of broth was investigated. pH of the extract-added broth was lower than control at the initial, but higher after 72 hours of preservation, which showed that when it added 0.1% and 0.3% of extract to the broth, pH of Kaempferia galanga L. was 4.92 and 5.08 respectively, whereas control was 4.60. Titratable acidity was lowered after 48 hours and also Kaempferia galanga L. showed the lowest acidity with 0.66 for adding 0.1% of its extract and 0.55 for 0.3% of adding, but control was 0.89 at the time of 90 hours of preservation, and then it showed to be lowered in the order of Zanthoxylum bungeanum, Eugenia caryophyllata and Foeniculum vuigare. Turbidity of each broth added the extracts of four of the medicinal plants was 7.5∼7.9 and 7.9∼8.2, respectively for 0.1% and 0.3% of concentration at the initial, but it began to lower and 90 hours later it was 8.8∼9.5 and 8.7∼9.0 respectively, whereas control was 10.8. Total viable cells(TVC) and coliform bacteria(CB) were increased with great at the 72 hours of preservation time, and Kaempferia galanga L. was the most effective, which when control was 4.8${\times}$10 CFU/ml at 72 hours, TVC was 1.7${\times}$10 CFU/ml for the addition of 0.1% of extract and 0.9${\times}$10 CFU/ml for 0.3%. CB was 3.2${\times}$10 CFU/ml for 0.1% and 1.7${\times}$10 CFU/ml for 0.3% respectively and 6.0 ${\times}$ 10 CFU/ml for control at the time of 72 hours, and it was lowered in the order of Zanthoxylum bungeanum, Eugenia caryophyllata and Foeniculum vulgare. Volatile basic nitrogen content detected that control was 2.67mg% at first, and then increased to 3.96mg% at 90 hours of preservation, but the broth added with the extract of Kaempferia galanga L. was 2.58mg% for 0.1% and 2.47mg% for 0.3% at the initial, and at 90 hours it was 3.64mg% and 3.33mg% respectively. The results of adding the extracts of four medicinal plants for the improvement of the preservation time of Naengmyeon broth, were that the most effective medicinal plant was Kaempferia galanga L. and the antimicrobial activity of the medicinal plant extracts for Naengmyeon broth was highly effective after 3 days of preservation time.

Extractive Nitrogenous Constituents in Commercial Saeujeot, a Salted and Fermented Shrimp (Acetes japonicus) (시판(市販) 새우젓의 함질소(含窒素) 엑스성분(成分)에 관(關)한 연구(硏究))

  • Park, Choon-Kyu;Kim, Woo-Jun;Kim, Kui-Shik;Park, Jung-Nim
    • Korean Journal of Food Science and Technology
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    • v.28 no.6
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    • pp.1135-1141
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    • 1996
  • In order to investigate the composition and the actual status of extractive nitrogenous compounds in saeujeot, a salted and fermented seawater shrimp(Acetes japonicus), the extract was analyzed separately into extractive nitrogen, free amino acids, oligpopeptides, nucleotides and related compounds, quaternary ammonium bases, and guanidino compounds, using specimens collected at fish markets from July in 1994 November in 1995. The extractive nitrogen of saeujeot was ranging from 430 mg to 528 mg, and the total of free amino acids in it was ranging from 1,509 mg to 2,131 mg, The extract was rich in free amino acids especially in lysine, glutamic acid, leucine, arginine, glycine, histidine, and glutamine, However, the content of most free amino acids was fluctuated from collection to collection. The total of nucleotides and related compounds in the extract was ranging from $1.38\;{\mu}mol\;to\;7.41\;{\mu}mol$, and the rage of fluctuation was essential identical with those of the extractive nitrogen and free amino acids. Homarine, trigonelline, ${\beta}-alaninebetaine$, and glycinebetaine were found in the saeujeot extract. Among them, homarine was the most abundant, ranging from 97 mg to 224 mg, but trigonelline, ${\beta}-alaninebetaine$, and glycinebetaine were very low. TMAO and TMA in the saeujeot extract were $13{\sim}80\;mg\;and\;12{\sim}280\;mg$, respectively. A small amount of creatine (less than 6 mg) and creatinine (less than 1 mg) was detected in all samples.

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Cold Storage and Quality Stability of Ascidian, Halocynthia roretzi (우렁쉥이 저온저장 및 품질안정성에 관한 연구)

  • Lee, Kang-Ho;Lee, Min-Ju;Jung, Byung-Chun;Hong, Byeong-Il;Cho, Ho-Sung;Lee, Dong-Ho;Jung, Woo-Jin
    • Korean Journal of Food Science and Technology
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    • v.26 no.4
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    • pp.382-388
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    • 1994
  • Because of its restrictive harvest from spring to summer, It is necessary to preserve raw ascidians, Halocynthia roretzi, for the purpose of processing regardless of season. We evaluated low Temperature tolerance of ascidian and conditions for cold storage to secure the quality of the stock. To retard the browning of meat rapidly occurred after sucking, ascidians were blanched for 10 seconds in 10% boiling salt solution or dipped for 60 seconds in 0.2% $NaHSO_3$ solution, respectively. The samples were stored in ice, at $-17^{\circ}C$ or $-35^{\circ}C$ for 85 days, respectively. Changes in VBN, glycogen, brown pigment formation, total carotenoid, nucleotides and their related compounds during the storage were determined, and sensory evaluation of quality was also practiced. VBN and brown pigment formation were rapidly increased. Glycogen was gradually decreased and then not detectable after 85 days in case of ice storage. Lipophilic brown pigment was higher than hydrophilic and rapidly increased during storage. The result of sensory evaluation showed that the ascidian treated in 0.2% $NaHSO_3$ was good for 85 days of storage at $-35^{\circ}C$ . Judging from the results of chemical experiment and sensory evaluation, the quality of ascidian treated in 0.2% $NaHSO_3$ and stored at $-35^{\circ}C$ was better than that of other samples.

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Comparison of Flavor Characteristics and Palatability of Beef Obtained from Various Breeds (품종별 쇠고기의 풍미특성과 기호성 비교)

  • Park, Hyung-Il;Lee, Moo-Ha;Chung, Myung-Sub
    • Korean Journal of Food Science and Technology
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    • v.26 no.5
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    • pp.500-506
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    • 1994
  • For comparison of beef quality, four kinds of beef (Korean native cattle beef, dairy cattle beef, imported beef, cross-bred beef) were investigated through tenderness, juiciness and flavor related components measurement and organoleptic tests. Flavor related chemical components such as NPN, IMP, free fatty acid and free amino acids were analyzed, water holding capacity, contents of hydroxyproline and intramuscular fat were measured for evaluation of beef tenderness. Instron was also used for measuring beef tenderness as an objective method. Triangle test and descriptive analysis test were conducted for comparison and evaluation of preference of various beef samples. In hardness analysis using Instron, imported and cross-bred beefs had higher value than that of Korean native cattle or dairy cattle beef. Water holding capacity and pH of Korean cattle beef was higher than that of others. The intramuscular fat content of Korean cattle beef was highest, so it was expected juicier than other beef. In flavor related compound analysis, NPN content of Korean native cattle beef was the lowest, which shows it spent the least time among sample meats after slaughter. IMP, hypoxantine and inosine were most abundant in Korean native cattle beef. In free amino acids analysis showed that the proportion of basic acid and aromatic acid content of Korean native cattle beef was highest, whereas that of sulfur containing amino acid of imported beef was highest. TBA value of Korean native cattle beef was the lowest, and analysis of fatty acid composition revealed that the proportion of unsaturated fatty acid of Korean native cattle beef was higher than imported and dairy cattle beef, but similar to cross-bred beef. Organoleptic test was performed by triangle test and descriptive analysis. In triangle test, most panelist could distinguish Korea native cattle beef from imported beef and cross-bred beef, imported beef from cross-bred beef. In descriptive analysis which relys on subjective standards of panelists, there was no difference among beef in aroma, flavor and tenderness except juiciness. Even though contents of non volatile flavor compounds in Korean native cattle beef were higher than those of other beef samples, there were no significant differences in subjective panel test. The results showed that Korean consumers do not have common standards for beef quality evaluation.

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Generation of a Mammalian Gene Expression Vector Using Bovine Viral Diarrhea Virus (Bovine Vira1 Diarrhea Virus를 이용한 포유동물세포 발현벡터의 개발)

  • 이영민
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.86-95
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    • 2002
  • As a result of genome projects, the research to elucidate the function of a protein of interest has recently been well-recognized. In order to facilitate functional genomics, a useful mammalian gene expression vector is required. Using an infectious CDNA clone of BVDV pNADLclns-, we have developed a mammalian gene expression vector. In this study, a replication-competent full-length infectious CDNA clone containing puremycin acetyltransferase (pac) gene (pNADLclns-/pac) was successfully generated. The viral RNA replication and viral protein NS3 synthesis were examined by detecting metabollically $^{32}P$-labelled genomic viral RNA and immunoblotting with a mouse anti-NS3 antibody. To generate viral replicon as an expression vector, we examine if the viral structural genes (C, E0, El, E2) are required for viral replication by deletion analysis. As a result, all of the structural proteins are dispensable for viral replication per se, but essential for infectious viral particle formation. Based on our deletion analysis, we have generated a replication-competent BVDV viral replicon (pNADLclns-/pac/${\Delta}S$), whose structural genes are all deleted. In addition to NADLclns- /pac/${\Delta}S$, NADLclns-/ luc/${\Delta}S$ viral replicon containing luciferase gene as a reporter was constructed and fecund to be replication-compotent in HeLa and BHK cells as well as MDBK cells. Therefore, BVDV viral replicon developed in our study will be a useful tool to express a protein of interest in various mammalian cells.

Primer Evaluation for the Detection of Toxigenic Microcystis by PCR (독소 생성 Microcystis 검출을 위한 PCR primer의 평가)

  • 이현경;김준호;유순애;안태석;김치경;이동훈
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.166-174
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    • 2003
  • Microcystin produced by cyanobacteria in surface waters, such as eutrophic lake and river, is a kind of serious environmental problems due to its toxicity to human and wild animals. Microcystin is synthesized nonribosomally by the large modular multi-functional enzyme complex known as microcystin synthetase encoded by the mcy gene cluster. Amplification of mcy genes by PCR from cultures and environmental samples is a simple and efficient method to detect the toxigenic Microcystis. In order to evaluate primers designed to detect toxic microcystin-producing strains, 17 cyanobacterial strains and 20 environmental samples were examined by PCR with 7 pairs of primers. Some microcystin-producing cyanobacteria were not detected with FAA-RAA, TOX4F-TOX4R and FP-RP primers. The fragment of unexpected size was amplified with NSZW2-NSZW1 primers in Microcystis strains isolated from the lakes in Korea. TOX1P-TOX1F primers failed in amplification of toxin-producing strains. Only MSF-MSR and TOX2P- TOX2F primers amplified the fragments of mcy genes from 11 strains of microcystin-producing Microcystis. The water samples taken from 20 lakes in Korea were analyzed by PCR using each of the primers. In all the water samples, cyanobacteria capable of producing microcystin were detected by the PCR with TOX2P-TOX2F primers. These results indicate that TOX2P-TOX2F primers are better than the other primers for detection of microcystin-producing Microcystis strains in Korea. The nucleotide sequences of mcy gene in Microcystis aeruginosa NIER10010 suggest genetic diversity of Korean isolates.

Regulatory Characterization of xylA Promoter Region in Escherichia coli (대장균의 xylA 프로모터 영역의 조절 특성)

  • Kang, Byung-Tae;Roh, Dong-Hyun;Joo, Gil-Jae;Rhee, In-Koo
    • Applied Biological Chemistry
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    • v.39 no.6
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    • pp.443-448
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    • 1996
  • In order to investigate the function of xylA promoter(Pxyl) as regulatory region Pxyl-lacZ fusion gene was constructed by the insertion of xylA promoter to the multiple cloning site of upstream of lacZ gene in a multicopy numbered plasmid pMC1403 containing promoterless lac operon, which was designated pMCX191, and Pxyl-lacZ fragment from pMCX191 was inserted to low copy numbered plasmid pLG339, designated pLGX191. The expressions of ${\beta}-galactosidase$ in these recombinant plasmids containing Pxyl-lacZ fusion gene were induced strongly by the addition of xylose, repressed by the addition of 0.2% glucose in the presence of xylose. The catabolite repressions were derepressed by the addition of 1 mM cAMP as same as native xylA gene. The fragment of xylA promoter was partially deleted from the upstream of xylA promoter by exonuclease III to investigate the regulation site of xylA promoter and the degrees of deletion derivatives of xylA promoter were analyzed by the DNA base sequencing. By the investigations of the induction by xylose, repression by glucose and derepression by cAMP on xylose isomerase production, the regulation site of xylA promoter may be located in segment between -165 and -59 bp upstream from the initiation site of xylA translation.

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