• Title/Summary/Keyword: Molecular length

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Molecular Cloning of the cDNA of Heat Shock Protein 88 Gene from the Entomopathogenic Fungus, Paecilomyces tenuipes Jocheon-1

  • Liu, Ya-Qi;Park, Nam Sook;Kim, Yong Gyun;Kim, Keun Ki;Park, Hyun Chul;Son, Hong Joo;Hong, Chang Ho;Lee, Sang Mong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.28 no.2
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    • pp.71-84
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    • 2014
  • The full-length heat shock protein 88 (HSP88) complementary DNA (cDNA) of Paecilomyces tenuipes Jocheon-1 was obtained by screening the Paecilomyces tenuipes (P. tenuipes) Jocheon-1 Uni-Zap cDNA library and performing 5' RACE polymerase chain reaction (PCR). The P. tenuipes Jocheon-1 HSP88 cDNA contained an open reading frame (ORF) of 2,139-basepair encoding 713 amino acid residues. The deduced amino acid sequence of the P. tenuipe s Jocheon-1 HSP88 cDNA showed 77% identity to Nectria haematococca HSP88 and 45-76% identity to other fungal homologous HSP88s. Phylogenetic analysis and BLAST program analysis confirmed that the deduced amino acid sequences of the P. tenuipes Jocheon-1 HSP88 gene belonged to the ascomycetes group within the fungal clade. The P. tenuipes Jocheon-1 HSP88 also contained the conserved ATPase domain at the N-terminal region. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as an 88 kilodalton (kDa) polypeptide in baculovirus-infected insect Sf9 cells. Under higher temperature conditions for the growth of the entomopathogenic fungus, mRNA expression of P. tenuipes Jocheon-1 HSP88 was quantified by real time PCR (qPCR). The results showed that heat shock stress induced a higher level of mRNA expression compared to normal growth conditions.

Enhanced Transdermal Delivery of Drug Compounds Using Scalable and Deformable Ethosomes (에토좀 입자크기와 멤브레인 특성 조절을 통한 약물의 경피흡수능 향상)

  • An, Eun-Jung;Shim, Jong-Won;Choi, Jang-Won;Kim, Jin-Woong;Park, Won-Seok;Kim, Han-Kon;Park, Ki-Dong;Han, Sung-Sik
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.2
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    • pp.105-113
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    • 2010
  • This study introduces a flexible approach to enhance skin permeation by using ethosomes with deformable lipid membranes as well as controllable sizes. To demonstrate this, a set of ethosomes encapsulating an anti-hair loss ingredient, Triaminodil$^{TM}$, as a model drug, were fabricated with varying their size, which was achieved by solely applying the different level of mechanical energy, while maintaining their chemical composition. After characterization of the ethosomes with dynamic light scattering, transmission electron microscopy, and deformability measurements, it was found that their membrane deformability depended on the particle size. Moreover, studies on in vitro skin permeation and murine anagen induction allowed us to figure out that the membrane deformability of ethosomes essentially affects delivery efficiency of Triaminodil$^{TM}$ through the skin. It was noticeable in our study that there existed an optimum particle size that can not only maximize the delivery of the drug through the skin, but also increase its actual dermatological activity. These findings offer a useful basis for understanding how ethosomes should be designed to improve delivery efficiency of encapsulated drugs therein in the aspects of changing their length scales and membrane properties.

Distributions of Hyperfine Parameters in Amorphous $Fe_{83}B_9Nb_7Cu_1$ Alloys (비정질 $Fe_{83}B_9Nb_7Cu_1$의 M$\)

  • 윤성현;김성백;김철성
    • Journal of the Korean Magnetics Society
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    • v.9 no.6
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    • pp.271-277
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    • 1999
  • Amorphous $Fe_{83}B_9Nb_7Cu_1$ alloy has been studied by M$\"{o}$ssbauer spectroscopy. Revised Vincze method was used and distributions of hyperfine field, isomer shift, and quadrupole line broadening of the sample at various temperatures have been evaluated and Curie temperature and $H_{hf}\;(0)$ were calculated to be 393 K and 231 kOe, respectively. Temperature variation of reduced average hyperfine field shows a flattered curvein comparison with the Brillouin curve for S=1. This behavior can be explained on the basis of Handrich molecular field model, in which the parameter Δ, which is a measure of fluctuation in exchange interactions, is assumed to have the temperature dependence ${Delta}=0.75-0.64{\tau}+0.47{\tau}^2$ where $\tau$ is $T/T_C$. At low temperature, the average hyperfine field can be fitted to $H_{hf}\;(T)=H_{hf}\;(0)\;[1-0.44\;(T/T_C)^{3/2}-0.28(T/T_C)^{5/2}-… ]$, which indicates the presence long wave length spin wave excitations. At temperature near TC, reduced average hyperfine field varies as $1.00\;[1-T/T_C]^{0.39}$. It is also found that half-width of the hyperfine field distribution was 102 kOe (3.29 mm/s) at 13 K and decreased monotonically as temperature increased. Above the Curie temperature, an average quadrupole splitting value of 0.43 mm/s was found. Average line broadening due to quadrupole splitting distribution was 0.31 mm/s at 13 K and decreases monotonically to 0.23 mm/s at 320 K, whereas that due to the isomer shift distribution is 0.1 mm/s at 13 K and 0.072 mm/s at 320 K, which is much smaller than that of both hyperfine field and quadrupole splitting. The temperature dependence of the isomer shift can be fitted within the harmonic approximation to a Deybe model with a Debye temperature ${Theta}_D=424{\pm}5K$.TEX>.

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Characterization of Somatolactin cDNA from Rock Bream (Oplegnathus fasciatus) (돌돔(Oplegnathus fasciatus) somatolactin cDNA의 분석)

  • 강현실;여인규;이제희
    • Journal of Life Science
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    • v.13 no.6
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    • pp.805-813
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    • 2003
  • cDNA encoding somatolactin (SL) was obtained by RT-PCR from pituitary glands of rock bream (Oplegnathus fasciatus). The full length cDNA of rock bream somatolactin (rbSL) is 1636 bp long. It contains a 696 bp open reading frame encoding a signal peptide of 24 amino acids (an) and a mature protein of 207 aa. rbSL has seven cysteine residues$(Cys^{5},\; Cys^{15},\; Cys^{42},\; Cys^{65},\; Cys^{181},\; Cys^{198}\; $and $Cys^{206})$ and two potential N-glycosylation sites at positions $Asn^{121}$and $Asn^{153}$. The rbSL shares 61.1∼92.6% amino acid sequence similarities and 63∼92.6% nucleotide sequence identities with other teleost SLs, except for goldfish and channel catfish SL. Amino acid sequence alignment revealed that rbSL has four conserved domains $(A_{SL},\; B_{SL},\; C_{SL}and\; D_{SL})$ common to all SLs. Out of these domains, $(A_{SL},\; B_{SL},\; C_{SL}and\; D_{SL})$, are also conserved in all teleost growth hormones and prolactins. The cDNA of rbSL has been cloned into pET expression vector in order to produce recombinant rbSL in E. coli BL2l(DE3) cells. The recombinant protein showed a molecular weight of 27 kDa in SDS-PAGE.

Pharmaco-medical Application of Antimicrobial Peptides Derived from Insect (곤충유래 항균 펩티드의 의약학적 적용)

  • Lee, Joon Ha;Kim, In-Woo;Kim, Mi-Ae;Yun, Eun Young;Hwang, Jae Sam
    • Journal of Life Science
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    • v.26 no.6
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    • pp.737-748
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    • 2016
  • By this time, insect antimicrobial peptides (AMPs) have been characterized more than 150 peptides since purification of cecropin in the hemolymph of pupae from Hyalophora cecropia in 1980. Therefore, it is considered that insects are good sources of AMP selection. Insect AMPs are small (low molecular weight) and cationic, and amphipathic with variable length, sequence, and structure. They perform a pivotal role on humoral immunity in the insect innate immune system against invading pathogens such as bacteria, fungi, parasites, and viruses. Most of the insect AMPs are induced rapidly in the fat bodies and other specific tissues of insects after septic injury or immune challenge. Then the AMPs subsequently released into the hemolymph to act against microorganisms. These peptides have a broad antimicrobial spectrum against various microbes including anticancer activities. Insect AMPs could be divided into four families based on their structures and sequences. That is the α-helical peptides, cysteine-rich peptides, proline-rich peptides, and glycine-rich peptides/proteins. For instance, cecropins, insect defensins, proline-rich peptides, and attacins are common insect AMPs, but gloverins and moricins have been identified only in lepidopteran species. This review focuses on AMPs from insects and discusses current knowledge and recent progress with potential applications of insect AMPs.

Molecular Cloning and Expression of the Metallothionein Gene under Environmental Stresses in Sweet Potato (고구마 metallothionein 유전자의 클로닝 및 환경 스트레스 하에서 발현 분석)

  • Kim, Young-Hwa;Yu, Eun Jeong;Huh, Gyung-Hye
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1415-1420
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    • 2017
  • The metallothionein (MT) gene (IbMT3) was selected from an EST library of suspension-cultured sweet potato cells. The MT gene, which is one of abundant ESTs in the library, is involved in stress regulation of cells and tissues. A full-length IbMT3 cDNA was obtained and analysis of its nucleotide sequence revealed that IbMT3 encoded a type 3 MT protein, based on its structural characteristics. The function of type 3 MT in plants is not yet known. Northern blot analysis showed stronger expression of IbMT3 in suspension-cultured cells than in sweet potato plant leaves. Since cell culture is known to impose a state of oxidative stress on cells, sweet potato plants were subjected to oxidative stress to investigate the transcriptional regulation of IbMT3. When the herbicide methyl viologen (MV) was administered for 6, 12, and 24 hr, IbMT3 transcription rapidly increased at 6 hr and then decreased. A cold treatment at $15^{\circ}C$ for 24 and 48 hr resulted in a gradual increase in IbMT3 expression. These findings indicate that IbMT3 expression is regulated in response to environmental and oxidative stress. IbMT3 isoform is expected to have antioxidant effects in sweet potato plants and may play an important role in cellular adaptation to oxidative stress.

Characterization of Streptococcosis Occurrence and Molecular Identification of the Pathogens of Cultured Flounder in Jeju Island (제주지역 양식넙치의 연쇄구균증 발생동향 및 원인균에 대한 분자적 동정)

  • Jeong, Yong-Uk;Kang, Chul-Young;Kim, Min-Ju;Heo, Moon-Soo;Oh, Duck-Chul;Kang, Bong-Jo
    • Korean Journal of Microbiology
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    • v.42 no.3
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    • pp.199-204
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    • 2006
  • Streptococcosis of olive flounder(Paralichthys olivaceus) is an important bacterial disease in Jeju island. In this study, we investigated monthly infection pattern of this disease in different size of the flounder fish. Even though the disease occurred throughout the year, the infection ratio was relatively higher in the months with warm water season. The infection was more prevalent in adult flounder over 30 cm total length compare to these of small size fish. Two infectious species of streptococcosis pathogens were detected by multiplex PCR assay. Detection ratios of Streptococcus iniae and S. parauberis reached up to 46% and 54%, respectively, from June 2003 to May 2005 in Jeju island. S. iniae occurred intensively from September to October, whereas S. parauberis reported from March to May. S. iniae and S. parauberis infections of cultured flounder share some common features, but clinical findings showed considerable differences between two diseases. Distended abdomen, protruded anus and ascitic fluid in the peritoneal cavity are evident lesions detected in S. iniae infection, whereas, flounders infected by S. parauberis showed prominent lesions such as darkened surface and haemorrhaging in the non-ocular side. Both streptococcosis pathogens were sensitive to antibiotics, such as ampicillin and amoxicillin. However, S. iniae strains were more sensitive to doxycycline, erythromycin and oxytetracycline than S. parauberis strains.

Fine Structural Analysis of Principal and Secondary Eyes in Wandering Spider, Pardosa astrigera (배회성 거미 (Pardosa astrigera) 주안과 부안의 미세구조적 분석)

  • Jeong, Moon-Jin;Lim, Do-Seon;Moon, Myung-Jin
    • Applied Microscopy
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    • v.30 no.1
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    • pp.1-9
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    • 2000
  • The wandering spider, Pardosa astrigera, had four pairs of ocelli that arranged in three rows on the cephalothorax. Along the anterior margin lay a pair of small anterior median (AM) eye flanked on each side by anterior lateral (AL) eye. Two large posterior median (PM) eye was situated on the clypeus behind the anterior row and still more posteriorly was a pair of posterior lateral (PL) eye. The visual cell of retina consisted of cell body, rhabdome, and intermediate segment. Bipolar neuron was found in anterior median eye (principal eye) and unipolar neuron in others (secondary eye). Rhabdome showed that arranged in PMeye and PLeye. But rhabdomes of AMeye and ALeye were irregular in retina. Except AMeye, incontinuous tapetum found in ALeye, PMeye, PLeye. Anterior median eye was similar to anterior lateral eye in length and posterior median eye similar to posterior lateral eye. Component size of eye were similar to 4 pairs eye in cornea. Size of lens, cell body, and rhabdome was similar not only anterior median eye and anterior lateral eye but also posterior median eye and posterior lateral eye. Vitreous body was large posterior median eye than others.

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PCR-RFLP for the Identification of Mammalian Livestock Animal Species

  • Han, Sang-Hyun;Park, Seon-Mi;Oh, Hong-Shik;Kang, Geunho;Park, Beom-Young;Ko, Moon-Suck;Cho, Sang-Rae;Kang, Yong-Jun;Kim, Sang-Geum;Cho, In-Cheol
    • Journal of Embryo Transfer
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    • v.28 no.4
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    • pp.355-360
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    • 2013
  • Precise, rapid and simple methods for species identification in animals are among the most important techniques in the livestock industry and research fields including meat classification. In this study, polymerase chain reaction (PCR) based molecular identification using inter species polymorphisms were examined by PCR-restriction fragment length polymorphism (RFLP) analysis for mitochondrial DNA (mtDNA) cytochrome b (CYTB) gene sequences among four mammalian livestock animals (cattle, horse, goat and pig). The results from PCR-RFLP analysis using the AluI restriction enzyme were also provided for the species-specific band patterns among CYTB gene sequences in these four species. The AluI-digestion for CYTB genes provided interesting migration patterns differentially displayed according to each species. Cattle and horse had one AluI-recognition site at different nucleotide positions and their AluI-digested fragments showed different band patterns on the gels. Pig had two AluI-recognition sites within the amplified CYTB sequences and produced three bands on the gels. Goat had no AluI-recognition site and was located at the same position as the uncut PCR product. The results showed the species-specific band patterns on a single gel among the four livestock animal species by AluI-RFLP. In addition, the results from blind tests for the meat samples collected from providers without any records showed the identical information on the species recorded by observing their phenotypes before slaughter. The application of this PCR-RFLP method can be useful and provide rapid, simple, and clear information regarding species identification for various tissue samples originating from tested livestock species.

Identification of Arbuscular Mycorrhizal Fungi Colonizing Panax ginseng Using 18S rDNA Sequence (18S rDNA를 이용한 인삼(Panax ginseng)의 내생균근 균의 동정)

  • Eo, Ju-Kyeong;Kim, Dong-Hun;Jeong, Hyeon-Suk;Eom, Ahn-Heum
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.182-186
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    • 2004
  • Morphological observation of roots and molecular technique were used to investigate the symbiotic relationships between arbuscular mycorrhizal (AM) fungi and ginseng roots. Korean ginseng, Panax ginseng, was collected from 8 sites in Korea. Colonization pattern of AM fungi in ginseng roots was determined as an Arum type under light microscopes. Nested PCR using AM fungal specific primers was employed to amplify a partial region on 18s rDNA of AM fungi from the root extracted mixed DNA. The amplified DNA was cloned and analyzed by random fragment length polymorphism (RFLP) with restriction enzymes, AluI, HinfI and AsuC21. One from each RFLP pattern was selected for sequencing. A total 16 clones were sequenced and identified as 2 species of AM fungi; Paraglomus brasilianum and Glomus spurcum. Paramglomus brasilianum was found from most of the ginseng roots, in this syudy suggesting that this species of AM fungi could have specific relationship with the ginseng root. Possible roles of AM fungal species in ginseng roots are discussed.