• Title/Summary/Keyword: ITS1 / 5.8S / ITS2

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Purification and N-Terminal Study of Bence Jones Proteins (Bence Jones 단백질(蛋白質)의 정제(精製) 및 N-말단검출(末端檢出))

  • Kim, Jun-Pyong
    • Applied Biological Chemistry
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    • v.13 no.1
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    • pp.59-64
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    • 1970
  • Human Bence Jones Protein could be purified by DEAF-Sephadex A-50 column $(2{\times}37cm)$ with 0.02M phosphate Buffer (pH 8.0) and gradient increasing with NaCl concentration as in Fig. 2-4. Sample As (K-type Bence Jones Protein) had two component, F-I was major component and its dried weight was 350mg. of starting material of 500mg. Other Sample Im and Ik (${\lambda}$-type Bence Jones Protein) was purified by DEAE-Sephadex A-50 with 0.02M phosphate Buffer(pH 8.0)too. F-I (major component) of Im and F-I of Ik were 242mg and 146mg. its dried weight respectively. K-type of Bence Jones Protein's(As, Ko, Ta.) N-terminal amino acid residue was determined by method of DNP,. K-type of Bence Jones Protein's amino acid residue were either glutamic acid or aspartic acid. Sample Ta was confirmed as glutamic acid its N-Terminal. As and Ko were aspartic acid. Each yellowish spot (DNP-amino acids) were extracted with 4ml. of pH 8.05% $NaHCO_3$ solution and calculated its recovery by O.D. $(360m{\mu}$ using the ${\varepsilon}=18.1{\times}10^3DNP$ $Asp\;{\varepsilon}=17.41{\times}10^(3)\;DNP\;Glu$ considering 50% lose during; the acid (6N-HCI) hydrolysis. Recovery of ko and As were 54.3% and 65% of its starting materials (DNP-Protein). Sample Ta's recovery was 85% of its DNP-protein. ${\lambda}$-type of Bence Jones Protein was rot investigated its N-terminal amino acid residue by DNP-method, probably it was blocked its N-terminal residue with glutamic acid.

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Phylogenetic Study of Genus Haliotis In Korea by Internal Transcribed Spacer Sequence (ITS) (ITS에 의한 한국내 전복 속 분류군의 유전적 계통분류학적 연구)

  • Huh, Man-Kyu;Kim, Jung-Ho;Moon, Du-Ho
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1003-1008
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    • 2009
  • Abalone (genus Haliotis) is a woody species with a long life span that is primarily distributed throughout the world, including Asia. This species is regarded as a very important marine gastropod mollusk in Korea and China, and also in food industries around the world. We evaluated a representative sample of the five species with nuclear ribosomal DNA internal transcribed spacer sequences (ITS) to estimate genetic relationships within the genus. Aligned nucleotide sequences of the length of the 5.8S subunit of all taxa of Haliotis were found to constant of 160 bp nucleotides. However, aligned nucleotide sequences of the length of ITS1 were varied within genus Haliotis, varying from 272 in H. diversicolor aquatilis to 292 in H. discus hannai. Aligned nucleotide sequences of the length of ITS2, especially, vary from 722 in H. diversicolor aquatilis to 752 in H. sieboldii. Total alignment length is 763 positions, of which 78 are parsimony-informative, 57 variable but parsimony-uninformative, and 459 constant characters. H. discus hannai was similar to H. discus, while H. diversicolor aquatilis was more distinct. ITS analysis may be useful in germ-plasm classification several taxa of genus Haliotis.

Phylogenetic Analysis of Agaricus blazei and Related Taxa by Comparing the Sequences of Internal Transcribed Spacers and 5.8S rDNA (Internal Transcribed Spacer와 5.8S ribosomal DNA의 염기서열 분석에 의한 Agaricus blazei와 근연종에 대한 계통분류학적인 연구)

  • 김기영;하명규;이태호;이재동
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.180-184
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    • 1999
  • Molecular spslemaucs of Agaricus species was investigated on the base of the sequences of the internal transcribed spaceriITS) regions in ribosomal DNA (rDNA). The sequences of the ITS region in 5 species and two group of Agaricus genus were resolved. In the phylogenetic trees. the species generally divided inlo two subclusters, refered to here as the group I and group 11. The group I consisted of Agaricus blazei ATCC 76739, Agarictrs blazei species cultivated in Korean hmings. Ago/-icus anmensis IMSNU 32049 and Agaricus can~pestris VPI-OKM 25665. Between Agaricus blazei NCC 76739 and the Agaricus blazei species cultivated in Korean farmings had the variation in lhe 5 nucleotide on the ITS regions. These varieties were presumed the variation by the geographic and cultivated conditions. In addition the subgroup of group I was formed by Agaricus arvensis LMSNU 32049 and Agaricus carnpests VPI-OKM 25665. The group IT included Agnrictrs bispoms CH 3004 and Agaricus pocillotor DUKE-J 173.

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Phylogenetic Status of an Undiscovered Zygomycete Species, Syncephalastrum monosporum, in Korea

  • Duong, Tham Thi;Nguyen, Thi Thuong Thuong;Jeon, Sun Jeong;Lee, Hyang Burm
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.371-376
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    • 2016
  • During a survey of undiscovered taxa in Korea, two zygomycete fungal isolates, EML-BT5-1 and EML-BT5-2, were isolated from the seed of a pumpkin (Cucurbita pepo) fruit in Korea. Based on their morphological characteristics and a sequence analysis of four genes, ITS1-5.8S-ITS2, 18S, 28S rDNA, and EF-$1{\alpha}$, the isolates were confirmed to be Syncephalastrum monosporum in the family Syncephalastraceae. To our knowledge, the zygomycete fungal species S. monosporum has not been previously described in Korea.

Phylogenesis of Halophila ovalis (R. Br.) Hook. fil. (Hydrocharitaceae) from An Island, Korea (전남 여수시 안도섬에서 발견된 해오말의 유전학적 관계 연구)

  • Kim, Jeong-Bae;Cho, Eun-Seob
    • Journal of Life Science
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    • v.18 no.6
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    • pp.759-763
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    • 2008
  • Halophila ovalis (R. Br.) Hook. fil. was first collected from the Ando, Yeosu, Korea in 2007. H. ovalis is Widely distributed from sub-tropical to even tropical areas and produces the seeds using bisexual reproduction. Its leaf shape was oblong to ovate. Its leaf blades were rigid in texture, with a strong support to the leaf. Erect shoots arose at irregular intervals along the rhizome. The distance between the intramarginal vein and leaves margin was small. Nucleotides in ITS 1 and ITS 2 regions between the Korean and Japanese H. ovalis were found to be 100% similar, whereas Korean H. ovalis was found to have four nucleotides in the positions of 202 bp to 206 bp for 5.8S. In the analysis of the phylogenetic relationship using NJ method, Korean H. ovalis had a monophyletic genetic tree with Japanese H. ovalis, but no phylogenetic relationship with types from the Philippines, Australia, Malaysia, and Vietnam. The first occurrence of H. ovalisin Korea was associated with a strong migration of gene flow from Japan and high water temperature caused by the variations in climate.

Identification of Phellinus linteus by Morphological Characteristics and Molecular Analysis (형태적.분자생물학적 방법에 의한 Phellinus linteus의 동정에 관한 연구)

  • Kim, Sang-Hee;Kim, Soo-Ho;Sung, Jae-Mo;Harrington, Thomas C.
    • The Korean Journal of Mycology
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    • v.27 no.5 s.92
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    • pp.337-340
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    • 1999
  • The context and upper surface of Phellinus basidiocarp become blackened, rimose and woody. The basidiocarp is sessile, dimidiate and elongate. The basidiospores are pigmented and ovoid to globose. Hymenial setae are $17{\sim}35{\times}6{\sim}8{\mu}m$. Nineteen isolates of Phellinus species, including Phellinus linteus, were used for sequencing of the internal transcribed spacer (ITS) region of the nuclear rDNA. Based on these sequence data, specific primers were designed for identification of Phellinus linteus isolates in Korea. The specific primers were within the ITS1 and ITS2 regions and were nested within the universal primers flanking the spacer regions. A total of four primers (the universal primers ITS-1F and ITS-4, and the specific primers PL-F and PL-R) were used for detection of Phellinus linteus collected in Korea. The length of the four amplification products of Phellinus linteus DNA were 800 bp (ITS-1F/ITS-4), two bands of about 720 bp (ITS-1F/PL-R and PL-F/ITS-4), and 610 bp (PL-F/PL-R). Among 23 isolates of Phellinus species collected in Korea, Thirteen isolates were identified as Phellinus linteus based on the presence of the four bands. The other species produced only the single ITS-1F/ITS-4 product.

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Phylogenetic Relationships of Coprinoid Taxa and an Agaric-like Gastroid Taxon Based on the Sequences of Internal Transcribed Spacer (ITS) Regions (ITS 영역의 염기서열을 이용한 먹물버섯류 및 주름버섯 유사 복균류와의 계통학적 유연관계)

  • Park, Dong-Suk;Go, Seung-Joo;Ryu, Jin-Chang
    • The Korean Journal of Mycology
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    • v.27 no.6 s.93
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    • pp.406-411
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    • 1999
  • The phylogenetic relationships of black-spored basidioid taxa (mainly coprinoid taxa) and an agaric-like gastroid taxon were studied. The sequences of internal transcribed spacer regions (ITS) partially including 17S, 25S and 5.8S from 14 species (Coprinus comatus, C. atramentarius, C. micaceus, C. cinereus, C. disseminatus, C. rhizophorus, C. radians, C. echinosporus, Psathyrella candolleana, Podaxis pistillaris, Conocybe lactea, Bolbitius demangei, Agaricus balzei, and Stropharia rugosoannulata) were compared. The reciprocal homologies of ITS sequences among these species were in the range of $38.7{\sim}77.2%$. Black-spored taxa were classified into four clusters. Cluster I comprised C. micaceus, C. radians, and C. disseminata. Cluster II is consisted of C. cinereus, C. echinosporus, C. rhizophorus, and C. atramentarius. On the other hand, C. comatus is in cluster III with Agaricus balzei and Podaxis pistillaris even though this species is belonging to the section Coprinus in morphological aspect. Psathyrelloid taxon is included in cluster II. The question of the origin of secotioid (agaric-like) fungi has been taken, though largely on theoritical suggestions as to whether gastroid taxa give rise to agarics with secotioid taxa as intermediates or whether secotioid taxa are evolutionary novelities arising from many distinct groups of agarics. In this relationship, it was shown that secotioid taxon evolved from within agaric species.

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Anticarcinogenic Effect of Ginseng Powders Depending on the Types and Ages using Yun's Anticarcinogenicity Test (I) (벤조피렌으로 유도된 마우스 폐선종에 대한 인삼분말의 연근별 항발암효과)

  • Yun, Taik-Koo;Lee, Yun-Sil
    • Journal of Ginseng Research
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    • v.18 no.2
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    • pp.89-94
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    • 1994
  • The authors have already shown that 6 year old red ginseng extract or its powder has remarkable anticarcinogenic effects. In this study, we further investigated whether fresh ginseng or white ginseng has similar anticarcinogenic effects and also if their anticarcinogenic effects are related to the types and ages of ginseng using Yun's anticarcinogenicity test (9 week medium term bioassay model). Dried fresh ginseng and red ginseng at 1.5, 3, 4, 5 and 6 years, and while ginseng at 3, 4, 5 and 6 years were used. The following results were obtained: 1) In the dried fresh ginseng treated groups, the incidence of lung adenoma induced by benzo(a)pyrene was 41.39) and its incidence was reduced to 31.2%, 30.0%, 31.3%, 30.7% and 27.8% after co-treatment with 1.5, 3, 4, 5 and 6 year-dried fresh ginseng, respectively. A significant effect was observed only in 6 Year-dried fresh ginseng. 2) In the white ginseng treated groups, the incidence of lung adenoma induced by benzo(a)pyrene was 45.0% and its incidence decreased to 41.3%, 38.0%, 31.6%, and 25.3% after co-treatment with 3, 4, 5 and 6 year-white ginseng, respectively. Five and 6 year-ginsengs showed significant inhibition of lung adenoma. 3) In the red ginseng treated groups, the incidence of lung adenoma induced by benzo(a) pyrene was 48.6% and its incidence diminished to 37.9%, 41.7%, 31.7%, 28.3% and 25.5% after co-treat-melt with 1.5, 3, 4, 5 and 6 year-red ginseng, respectively. In 4, 5 and 6 year-ginsengs, the anticarcinogenic effect was prominent. From the above results, we concluded that a significant anticarcinogenic effect was observed in 6 year-dried fresh ginseng, 5 and 6 year-white ginsengs, and 4, 5 and 6 year-red ginsengs.

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Localization of Angiotensin II in Korean Bovine Follicles and Its Effects on IVM/IVF of Oocytes

  • Quen, J. H.;Kim, S. K.
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.252-252
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    • 2004
  • 1. The concentrations of Ang. Ⅱ were 7.2±0.91 × 10³, 3.8±0.34 × 10³, 3.5±0.30 × 10³, 2.8±0.22 × 10³ pg/㎖ in bovine follicular fluids from 1∼3 ㎜, 3∼5 ㎜, 5∼7 ㎜ and 8∼10 ㎜ follicles, respectively. However, the concentrations of Ang. Ⅱ decreased in follicular fluids from large follicles. (omitted)

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Isolation of a New Agar Degrading Bacterium, Maribacter sp. SH-1 and Characterization of its Agarase (신규 한천분해세균 Maribacter sp. SH-1의 분리 및 효소 특성조사)

  • Lee, Chang-Eun;Lee, Sol-Ji;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.156-162
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    • 2016
  • In this study, we isolated a new agar-degrading marine bacterium and characterized its agarase. An agardegrading marine bacterium SH-1 was isolated from seawater, collected from the seashore of Namhae in Gyeongnam province, Korea, and cultured in marine agar 2216 media. It was identified as Maribacter. sp. SH-1 by phylogenetic analyses, based on 16S rRNA gene sequence. The extracellular agarase was extracted from culture media of Maribacter sp. SH-1 and characterized. Its relative activities were 56, 62, 94, 100, and 8% at 20, 30, 40, 50, and 60℃, respectively, whereas 15, 100, 60, and 21% relative activities were observed at pH 5, 6, 7, and 8, respectively. Its extracellular agarase exhibited maximum activity (231 units/l) at pH 6.0 and 50℃, in 20 mM Tris-HCl buffer. Therefore, this agarase would be applicable as it showed the maximum activity at the temperature at which the agar is in a sol state. Furthermore, the agarase activities remained over 90% at 20, 30, and 40℃ after 0.5 h exposure at these temperatures. Thin layer chromatography analysis suggested that Maribacter sp. SH-1 produces extracellular β-agarase, as it hydrolyzes agarose to produce neoagarooligosaccharides, such as neoagarohexaose (34.8%), neoagarotetraose (52.2%), and neoagarobiose (13.0%). Maribacter sp. SH-1 and its β-agarase would be useful for the production of neoagarooligosaccharides, which shows functional properties, like skin moisturizing, skin whitening, inhibition of bacterial growth, and delay in starch degradation.