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Extracellular Production of Alpha-Interferon by Recombinant Escherichia coli : Part I. Construction of Expression Vectors (유전자 재조합 대장균을 사용한 Alpha-interferon의 생산과 분비: 제 1 부. 발현벡터의 제작)

  • 노갑수;최차용
    • KSBB Journal
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    • v.5 no.1
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    • pp.49-58
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    • 1990
  • We constructed hybrid plasmids to allow controlled and extracellular production of human alpha-interferon in Escherichia coli. The hybrid plassmids were constructed by transferring alpha-lFN gene from plasmid Hif-2h which has the alpha-lFN gene at PstI restriction site of pBR322, to plasmids pIN -IIIB3 and pIN-IIIC3 at restriction sites between HindIII and BamHI. Plasmids pIN-IIIB3 and pIN-IIIC3 carry E. coli lipoprotein promoter, lac promoter and operator in tandem. The plasmids also have lacl genes which encode for lac repressors, which allows controlled expression of genes cloned to the plasmids by using of inducer IPTG. Lipoprotein signal sequence is located just ahead of cloning sites of the plasmids, which helps cells to excrete or secrete cloned gene products. Plasmid pUC9 was used as a intermediate vector for transferring of alpha-lFN gene from Hif-2h to pIN vectors in order to solve the problem of different restriction sites between Hif-2h and pIN vectors.

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Studies of the egg drop laying diseases from the mass zone layer (산란계 밀집지역의 산란저하성 질병에 관한 연구)

  • Lee Jeoung-Won;Eum Sung-Shim;Park In-Gyu;Bea Joung-Jun;Joung Dong-Suk;Song Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.28 no.2
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    • pp.121-146
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    • 2005
  • Newcastle disease (ND), infectious bronchitis (IB), low pathogenic avian Influenza (LPAI) and fowl typhoid (FT) have been known as egg drop laying diseases because of the serious layer damage from mass zone layer. In this study, such egg drop laying diseases were investigated. To access this study, we peformed to evaluate antibody titers in serum and isolated bacteria and virus from organs and feces on May, July and September in 2003. The distribution of ND from January to May, IB and LPAI from October to February of the next year, and FT from March to September were inspected by the question survey in 21 farms. ND revealed to be positive rates of 490 to 474 $(96.7\%)$ in May, 510 to 506 $(99.2\%)$ in July and 510 to 510 $(100\%)$ in September with hemagglutination inhibition (HI) test. The mean antibody titers were 10.2, 9.9 and 10.2, respectively. With regard to IB, 484 out of 490 samples $(98.7\%)$ in May, 508 of 510 $(99.6\%)$ in July and 509 of 510 $(99.8\%)$ in September showed positive results and the mean antibody titers were gradually increased with 8.2, 9.0 and 9.4, respectively. According to HI test of LPAI, the positive results were shown in 442 of 480 $(92.1\%)$, 394 of 494 $(79.8\%)$ and 402 of 483 $(83.2\%)$ in May, July and September, respectively The mean antibody titers were decreased with 4.6, 4.3 and 4.0. The distribution of LPAI also elicited the positive rates of 480 to 475 $(99.0\%)$ in May, 494 to 485$(98.2\%)$ in July, 483 to 472 $(97.7\%)$ in September as determined by ELISA and the mean S/P ratio were 2.319, 2.557 and 2.380, respectively. Compared ELISA results with HI test of LPAI the positive results were 480 to 422 $(92.1\%),\;475(99.0\%),\;494\;to\;394 (79.8\%),\;485 (98.2\%)\;and\;483\;to\;402(83.2\%),\;472(97.7\%)$. Therefore, the positive rate determined by ELISA was higher than that of HI test with 6.9, 18.4 and $14.5\%$, respectively. When performed RT-PCR for ND using organ and feces samples, the pathotypes were detected $5(15.6\%)\;in\;May,\; 2(5.3\%) in\;July,\;2(7.1\%)$ in September but there is no samples showing positive band for LPAI. In attempt to isolate Salmonella gallinarum, bacteria were obtained from 4 cases (12.5%) in May, 9 (23.6%) in July, 5 (17.8%) in September. Thus the highest rate for isolation revealed to be shown in July When evaluated the antimicrobial susceptibility to 18 isolated strains of 5. gallinarum, bacteria were sensitive to trimethoprim/sulfamethox$(61.1\%),\;kanamycin\;(55.5\%),\;ampicillin\;(55.5\%)$ and amoxacillin/clavulanic acid $(55.5\%)$, cephalothin $(50.0\%)$, but resistant to penicillin $(88.9\%)$, streptomycin $(88.9\%)$, erythromycin $(83_4\%)$ and tetracycline $(61.1\%)$. According to HI test of ND and LPAI using captured 164 wild Korean tree sparrows (Passer nontanus), the positive rates were $47.6\%\;and\;57.3\%$, and the mean HI titers were 5.32 and 4.02, respectively. 71 $(43.2\%)\;and\;58(35.3\%)$ in captured sparrows also showed more than 4 titers for HI test to ND and LPAI, respectively However, the attempt for isolation of viruses failed in all samples.

Characterization of ptsHI Operon from Leuconostoc mesenteroides SY1, a Strain Isolated from Kimchi

  • Park Jae-Yong;Jeong Seon-Ju;Chun Ji-Yeon;Lee Jong-Hoon;Chung Dae-Kyun;Kim Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.988-992
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    • 2006
  • The ptsHI operon from Leuconostoc mesenteroides ssp. mesenteroides SY1 (L. mesenteroides SY1), a strain isolated from kimchi, was cloned and characterized. The ptsH open reading frame (ORF) was 273 bp in size, which can encode a protein of 90 amino acid residues with a molecular weight of 9,212 Da. The pfsI ORF was 1,719 bp in size, which was capable of encoding a protein of 572 amino acids with a molecular mass of 62,549 Da. ptsH and pfsI genes were transcribed as a single transcript of 2.0 kb in size regardless of carbon sources, supporting the operon structure. Although the deduced amino acid sequences of the HPr and EI were highly homologous with those of other Gram-positive bacteria, an additional amino acid (glutamine at the $3^{rd}$ amino acid) was present in HPr from L. mesenteroides SY1. Phosphorylation sites of HPr included the histidine residue ($16^{th}$) and serine residue ($47^{th}$). Mutant HPrs, in which each phosphorylation site was mutated into alanine, were obtained, and phosphorylation with HPr and mutated HPrs showed that HPr was phosphorylated at the serine residue ($47^{th}$) by HPr kinaseiphosphorylase (HPr K/P).

A STUDY ON THE BOND STRENGTH OF RELINE RESIN TO PRESSURE INJECTION TYPE THERMOPLASTIC DENTURE BASE RESIN (가압주사식 열가소성 의치상 레진과 이장 레진 간의 결합강도에 관한 연구)

  • Moon Tae-Sung;Jeong Chang-Mo;Jeon Young-Chan;Lim Jang-Seop
    • The Journal of Korean Academy of Prosthodontics
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    • v.40 no.1
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    • pp.42-52
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    • 2002
  • The purpose of this study was to evaluate the bond strength of reline resin to pressure injection type thermoplastic denture base resin. The denture base resins used in this study were $Hi-polycarbonate^{(R)}$(High Dental Co., Japan), Acetal $dental^{(R)}$(Pressingdental s.r.1., Repubblica di San Marine) of thermoplastic resin and Acron $MC^{(R)}$(GC Dental Industrial Co., Japan) of heat cured resin. The reline resins used were Lucitone $199^{(R)}$(Dentsply international Inc., USA), Tokuso $rebase^{(R)}$(Tokuyama Corp., Japan), and $Lightdon-U^{(R)}$(Dreve-Dentamid-Gmbh, Germany). The reline resins are representative of heat-cured, self-cured, and light-cured resin respectively Bond strength was examined by use of a three-point transverse flexural strength test. The results were as follows 1. The bond strength of Lucitone 199 to Acron MC was the highest. 2. The bond strengths of Lucitone 199 and Tokuso rebase to Hi-polycarbonate resulted in a value of approximately one half that of Lucitone 199 to Acron MC and there were no significant differences between these and the bond strength of Tokuso rebase to Acron MC(p<0.05) 3. The bond strengths of reline resins to Acetal dental were lower than those of reline resins to Hi-polycarbonate. 4. For all base resins Lightdon-U showed lower bond strength than the other reline resins.

p53 Protein Expression in Imprint Cytology of Breast Carcinoma (유방암의 Imprint 표본에서 p53 단백 발현)

  • Kim, Dong-Sug;Lee, Eun-Hi;Kim, Ki-Kwon;Kim, Mi-Jin;Lee, Soo-Jung
    • The Korean Journal of Cytopathology
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    • v.6 no.1
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    • pp.1-9
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    • 1995
  • This study was carried out to determine the usefulness of imprint cytology for detecting p53 protein in breast carcinoma. NCL-DO7 (Novocastra, U.K.) was used to detect p53 protein immunocytochemically. A total of 33 cases was studied, Immunostaining of imprint cytology with NCL-DO7 was positive in 64% (21/33) and showed relatively high coincident rate (80%) with immunostaining of formalin-fixed, paraffin-embedded specimen p53 protein was related to negative estrogen receptor status, but not to the nuclear grade, lymph node metastasis, or tumor size. The fact that p53 protein expression was not related to nuclear grade might be due to predominance of nuclear grade 3. It was easier to determine the nuclear grade is one of the most important prognostic factors, in imprint cytology than in tissue specimen. p53 protein tended to be stained more strongly in imprint cytology than in tissue. It is concluded that the application of imprint cytology in p53 protein detection can be performed easily, and that it may contribute to the evaluation of prognostic factors in breast carcinoma.

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Subcloning and Sequencing of Maize rbcL Promoter Region

  • Woong-Seop Sim
    • Journal of Plant Biology
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    • v.38 no.1
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    • pp.107-113
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    • 1995
  • pRLYS1 containing intact rbcL gene of maize (Zea mays L. cv Golden X Bantam T-51; Zm-A) was digested with several restriction enzymes to construct subcones carrying promoter region of rbcL. The DNA fragments of 0.20, 0.19, 0.92 and 1.55 kb among the EcoRI digests, the EcoRI-DdeI digests, the AvaI digests and the EcoRI-BamHI digests of pRLYS1 were subcloned into pBluscriptSK+and named pRLPS2, pRLPS3, pRLPS14 and pRLPS35, respectively. Four subclones contain the 1.92 kb portion from 136 nucleotide downstream to 1780 nucleotide upstream from the ATG initiation codon of rbcL gene. pRLPS2 (-29 to -229) and pRLPS3 (-239 to -420 from the ATG) were sequenced. When nucleotide sequence of Zm-A was compared with sequence of rbcL promoter region of a different cultivar of maize (Zea mays L. cv WFG TMS X BS7; Zm-B), the difference rate between two cultivars was 4.3%. The mean of sequence divergence between Zm-A and three grass species in the same tribe, Andropogoneae, in the upstream region from 29 to 420 of ATG was 1.8%, whereas between Zm-B and above-mentioned three species was 5.4%. Therefore, Zm-A seems to evolutionarily closer to three other species in Andropogoneae tribe than Zm-B is.

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Wheel of Wellness Counseling in Community Dwelling, Korean Elders: A Randomized, Controlled Trial

  • Kwon, So-Hi
    • Journal of Korean Academy of Nursing
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    • v.45 no.3
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    • pp.459-468
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    • 2015
  • Purpose: The purpose of this study was to investigate the effects of Wheel of Wellness counseling on wellness lifestyle, depression, and health-related quality of life in community dwelling elderly people. Methods: A parallel, randomized controlled, open label, trial was conducted. Ninety-three elderly people in a senior welfare center were randomly assigned to two groups: 1) A Wheel of Wellness counseling intervention group (n=49) and 2) a no-treatment control group (n=44). Wheel of Wellness counseling consisted of structured, individual counseling based on the Wheel of Wellness model and provided once a week for four weeks. Wellness lifestyle, depression, and health-related quality of life were assessed pre-and post-test in both groups. Results: Data from 89 participants were analyzed. For participants in the experimental group, there was a significant improvement on all of the wellness-lifestyle subtasks except realistic beliefs. Perceived wellness and depression significantly improved after the in the experimental group (n=43) compared to the control group (n=46) from pre- to post-test in the areas of sense of control (p =.033), nutrition (p =.017), exercise (p =.039), self-care (p <.001), stress management (p =.017), work (p =.011), perceived wellness (p =.019), and depression (p =.031). One participant in the intervention group discontinued the intervention due to hospitalization and three in the control group discontinued the sessions. Conclusions: Wheel of Wellness counseling was beneficial in enhancing wellness for the community-dwelling elderly people. Research into long-term effects of the intervention and health outcomes is recommended.

ON THE STABILITY OF A CUBIC FUNCTIONAL EQUATION

  • Jun, Kil-Woung;Lee, Yang-Hi
    • Journal of the Chungcheong Mathematical Society
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    • v.21 no.3
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    • pp.377-384
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    • 2008
  • In this paper, we prove the stability of the functional equation $$\sum\limits_{i=0}^{3}3Ci(-1)^{3-i}f(ix+y)-3!f(x)=0$$ in the sense of P. $G{\breve{a}}vruta$ on the punctured domain. Also, we investigate the superstability of the functional equation.

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ON THE STABILITY OF A CAUCHY-JENSEN FUNCTIONAL EQUATION III

  • Jun, Kil-Woung;Lee, Yang-Hi;Son, Ji-Ae
    • Korean Journal of Mathematics
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    • v.16 no.2
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    • pp.205-214
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    • 2008
  • In this paper, we prove the generalized Hyers-Ulam stability of a Cauchy-Jensen functional equation $2f(x+y,\frac{z+w}{2})=f(x,z)+f(x,w)+f(y,z)+f(y,w)$ in the spirit of $P.G{\breve{a}}vruta$.

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WEIGHTED LEBESGUE NORM INEQUALITIES FOR CERTAIN CLASSES OF OPERATORS

  • Song, Hi Ja
    • Korean Journal of Mathematics
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    • v.14 no.2
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    • pp.137-160
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    • 2006
  • We describe the weight functions for which Hardy's inequality of nonincreasing functions is satisfied. Further we characterize the pairs of weight functions $(w,v)$ for which the Laplace transform $\mathcal{L}f(x)={\int}^{\infty}_0e^{-xy}f(y)dy$, with monotone function $f$, is bounded from the weighted Lebesgue space $L^p(w)$ to $L^q(v)$.

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