• Title/Summary/Keyword: P.C strand

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Backbone 1H, 15N, and 13C Resonance Assignments and Secondary-Structure of the Conserved Hypothetical Protein HP0892 of Helicobacter pylori

  • Han, Kyung-Doo;Park, Sung-Jean;Jang, Sun-Bok;Lee, Bong-Jin
    • Molecules and Cells
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    • v.25 no.1
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    • pp.138-141
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    • 2008
  • HP0892 (SwissProt/TrEMBL ID O25552) is a 90-residue conserved hypothetical protein from Helicobacter pylori strain 26695, with a calculated pI of 9.38 and a molecular mass of 10.41 kDa. It belongs to the Plasmid stabilization system protein family (PF05016) in the Pfam database. Proteins with sequence similarity to HP0892 exist in Vibrio choierae, Enterococcus faecalis, Campylobacter jejuni, Streptococcus pneumoniae, Haemophilus influenzae, Escherichia coli O157. Here we report the sequence-specific backbone resonance assignments of HP0892 using multidimentional heteronuclear NMR spectroscopy. About 97.0% (422/435) of the HN, N, CO, $C{\beta}$, $C{\alpha}$ resonances of 90 residues of HP0892 were assigned. On the basis of the resonance assignments, three helical regions and four strand regions were identified using the CSI program. This study is a prerequisite for calculating the solution structure of HP0892, and will be useful for studying its interaction with other molecules.

Mutational Analysis of the MTHFR Gene in Breast Cancer Patients of Pakistani Population

  • Akram, Muhammad;Malik, Fa;Kayani, Mahmood Akhtar
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1599-1603
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    • 2012
  • Objectives: Since methylenetetrahydrofolate reductase (MTHFR) maintains the balance of circulating folate and methionine and blocks the formation of homocysteine, its regulation in relation to different cancers has extensively been studied in different populations. However, information on Pakistani breast cancer patients is lacking. The MTHFR gene has two most common mutations that are single nucleotide additions which result in change of amino acids C677T to Ala222val and A1298C to Glu429Ala. Methodology: 110 sporadic breast patients with no prior family history of cancer or any other type of genetic disorders along with 110 normal individuals were screened for mutations in exons 1 to exon 9 using single strand conformational polymorphism, RFLP and sequencing analyzer. Results: The p values for the 677CC, 677CT, and 677TT genotypes were 0.223, 0.006, and 0.077, respectively. Those for the 1298AA, 1298AC, and 1298CC genotypes were 0.555, 0.009, and 0.003, respectively. Conclusions: We found an overall a significant, weak inverse association between breast cancer risk and the 677TT genotype and an inverse association with the 1298C variant. These results for MTHFR polymorphism might be population specific in sporadic breast cancer affected patients but many other factors need to be excluded before making final conclusions including folate intake, population and disease heterogeneity.

Nonstructural Protein 5B of Hepatitis C Virus

  • Lee, Jong-Ho;Nam, In Young;Myung, Heejoon
    • Molecules and Cells
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    • v.21 no.3
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    • pp.330-336
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    • 2006
  • Since its identification in 1989, hepatitis C virus has been the subject of extensive research. The biology of the virus and the development of antiviral drugs are closely related. The RNA polymerase activity of nonstructural protein 5B was first demonstrated in 1996. NS5B is believed to localize to the perinuclear region, forming a replicase complex with other viral proteins. It has a typical polymerase structure with thumb, palm, and finger domains encircling the active site. A de novo replication initiation mechanism has been suggested. To date, many small molecule inhibitors are known including nucleoside analogues, non-nucleoside analogues, and pyrophosphate mimics. NS5B interacts with other viral proteins such as core, NS3, 4A, 4B, and 5A. The helicase activity of NS3 seems necessary for RNA strand unwinding during replication, with other nonstructural proteins performing modulatory roles. Cellular proteins interacting with NS5B include VAMP-associated proteins, heIF4AII, hPLIC1, nucleolin, PRK2, ${\alpha}$-actinin, and p68 helicase. The interactions of NS5B with these proteins might play roles in cellular trafficking, signal transduction, and RNA polymerization, as well as the regulation of replication/translation processes.

Backbone 1H, 15N, and 13C Resonance Assignments and Secondary-Structure of Conserved Hypothetical Protein HP0894 from Helicobacter pylori

  • Han, Kyung-Doo;Park, Sung-Jean;Lee, Bong-Jin
    • Molecules and Cells
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    • v.20 no.3
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    • pp.442-445
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    • 2005
  • HP0894 (SwissProt/TrEMBL ID O25554) is an 88-residue conserved hypothetical protein from Helicobacter pylori strain 26695 with a calculated pI of 8.5 and a molecular weight of 10.38 kDa. Proteins with sequence similarity to HP0894 exist in Vibrio choierae, Enterococcus faecalis, Campylobacter jejuni, Streptococcus pneumoniae, Haemophilus influenzae, Escherichia coli O157, etc. Here we report the sequence-specific backbone resonance assignments of HP0894. About 97.5% (418/429) of the HN, N, CO, $C{\alpha}$, $C{\beta}$ resonances of the 88 residues of HP0894 were assigned. On the basis of these assignments, three helical regions and four strand regions were identified using the CSI program. This study is a prerequisite for calculating the solution structure of HP0894, and studying its interaction with its substrates, if any, and/or with other proteins.

Purification and Characterization of a Deoxyriboendonuclease from Mycobacterium smegmatis

  • Mandal, Prajna;Chakraborty, Phulghuri;Sau, Subrata;Mandal, Nitai Chandra
    • BMB Reports
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    • v.39 no.2
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    • pp.140-144
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    • 2006
  • A deoxyriboendonuclease has been purified to near homogeneity from a fast growing mycobacterium species, M. smegmatis and characterized to some extent. The size of enzyme is about 43 kDa as determined by a denaturing gel analysis. It shows optimum activity at $32^{\circ}C$ in Tris-HCl buffer (pH 7.2) containing 2.5 mM of $MgCl_2$. Both EDTA and $K^+$ but not $Na^+$ inhibit its activity. Evidences show that the enzyme is not a restriction endonuclease but catalyzes the endonucleolytic cleavage of both the double- as well as the single-strand DNA non-specifically. It has been shown that the cleavage by this enzyme generates DNA fragments carrying phosphate groups at 5' ends and hydroxyl group at the 3' ends, respectively. Analysis reveals that no endonuclease having size and property identical to our deoxyriboendonuclease had been purified from M. smegmatis before. The property of our enzymes closely matches with the deoxyriboendonucleases purified from diverse sources including bacteria.

Cytogenetic and Histological Characteristics of Ginseng Hairy Root Transformed by Agrobacterium rhizogenes (Agrobacterium rhizogenes에 의하여 형질전환된 인삼 모상근의 세포 유전학적 및 조직학적 특성)

  • 고경민
    • Journal of Plant Biology
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    • v.36 no.1
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    • pp.75-81
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    • 1993
  • Ginseng (Panax ginseng C.A. Meyer) hairy root transformed with Agrobacterium rhizogenes (strain A4) was examined cytogenetically and histologically to assess its characteristics. The optimum growth of hairy root obtained in hormone-free MS medium (sucrose 30 g/L, pH5-6) for long period cultures. All hairy root strains (HB1, HB2, HB3) had the 2n diploid number (2n=48) of chromosomes in root tip cells. There were no alteration in chromosome structure except in one cell of HB3 strain. Results of SDS-PAGE showed a few difference in pattern and number of bands between normal and hairy root of ginseng. The root anatomy of normal root and hairy root differed each other. The hairy root had a clearly defined vascular strand, and the morphology of cortical cells were disorganised with large intercellular spaces.

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Ethanol changes atpB gene expression and proton permeability in Streptococcus mutans (에탄올이 Streptococcus mutans의 atpB 유전자 발현 및 양성자 투과성에 미치는 영향)

  • Cho, Chul Min;Park, Yong Jin;Lee, Sae A;Kim, Jin Bom;Kang, Jung Sook
    • Journal of Korean Academy of Oral Health
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    • v.42 no.4
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    • pp.224-228
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    • 2018
  • Objectives: As a first step to study the anticaries effect of ethanol alone, we investigated the effects of ethanol on the expression levels of the atpB gene and proton permeability of Streptococcus mutans in suspension cultures. Methods: S. mutans UA159 was grown in brain heart infusion medium at either pH 4.8 or 6.8. The total extracted RNA was reverse-transcribed into cDNA using a $Superscript^{TM}$ First-Strand Synthesis System. The resulting cDNA and negative controls were amplified by ABI PRISM 7700 real-time PCR system with SYBR Green PCR Master Mix. For proton flux assay, bacterial suspensions were titrated to pH 4.6 with 0.5 M HCl, and then additional 0.5 M HCl was added to decrease the pH values by approximately 0.4 units. The subsequent increase in pH was monitored using a glass electrode. Ten percent (v/v) butanol was added to the suspensions at 80 min to disrupt the cell membrane. Results: In a concentration-dependent manner, ethanol alone not only decreased the growth rate of S. mutans and the expression of the atpB gene but also increased the proton permeability at both pH 4.8 and 6.8. Conclusions: These findings suggest that ethanol has the potential for an anticaries ingredient. We believe that ethanol may be used together with fluoride and/or other cariostatic agents in order to develop better anticaries toothpastes and/or mouthrinses.

Single Nucleotide Polymorphisms of the GnRHR Gene Associated with Reproductive Traits of Japanese Flounder (Paralichthys olivaceus)

  • He, Feng;Wen, Hai-Shen;Li, Ji-Fang;Yu, Da-Hui;Ma, Rui-Qin;Shi, Dan;Mu, Wei-Jie;Zhang, Yuan-Qing;Hu, Jian;Liu, Miao;Han, Wei-Guo;Zhang, Jia-Nan;Wang, Qing-Qing;Yuan, Yu-Ren;Liu, Qun
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.4
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    • pp.463-470
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    • 2011
  • Gonadotropin-releasing hormone receptor (GnRHR) gene is expressed at the anterior pituitary gland and plays a key role in gonad development. This study aimed to investigate molecular genetic characteristics of the GnRHR gene and elucidate the effects of single nucleotide polymorphisms (SNPs) of GnRHR gene on sex steroid level in Japanese flounder (Paralichthys olivaceus). We used polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and sequencing of the GnRHR gene in 75 individuals. We identified three SNPs in the GnRHR gene: P1 locus (C759A and C830T) in the coding region of exon2 which were both linked together and P2 locus (G984T) in the coding region of exon3, which added a new transcript factor (ADR1) and a new methylation site (CG). Only C830T of P1 leads to amino acid changes Thr266Ile. Statistical analysis showed that P1 was significantly associated with $17{\beta}$-estradiol ($E_2$) level (p<0.01) and gonadosomatic index (GSI) (p<0.05). Individuals with genotype BB of P1 had significantly higher serum $E_2$ levels (p<0.01) and GSI (p<0.05) than those of genotype AA or AB. Another SNP, P2, synonymous mutation, was significantly associated with GSI (p<0.05). Individuals with genotype AB of P2 had significantly higher GSI (p<0.05) than that of genotype AA. In addition, there was a significant association between one diplotype based on three SNPs and reproductive traits. The genetic effects for both serum $E_2$ level and GSI of diplotype D4 were super diplotypes (p<0.05). These results suggest that the SNPs in Japanese Flounder GnRHR are associated with $E_2$ level and GSI.

Studies on Cultural Characteristics of Ganoderma lucidum (Fr.) Karst (Ganoderma lucidum(Fr.) Karst의 배양적(培養的) 특성(特性)에 관(關)한 연구(硏究))

  • Seo, Geon Sik;Shin, Gwan Chull;Park, Jong Seong
    • Korean Journal of Agricultural Science
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    • v.15 no.1
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    • pp.27-35
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    • 1988
  • In order to elucidate the systematic taxonomy and genetic characters of Canoderma lucidum, cultural characteristics of the fungus were investigated. Mycelial growth of Ganoderma lucidum were favorable on oat meal agar medium, and optimum temperature and pH of the medium for mycelial growth were $30^{\circ}C$ and 5.5-6.0 respectively. Irradiation of white fluorescent lamp inhibited mycelial growth and critical time for inhibition of mycelial growth was 4-8 hours. Concentric zones and mycelial strands of Ganoderma lucidum was induced by irradiation of white fluorescent lamp and formation of mycelial sectors was influenced by nutrient source of media and irradiation of white fluorescent lamp. These characters were different among the isolates, but no relationship was observed between these characters and the fruiting body type of the fungus. Basidiospores were formed directly from the mycelium cultured on artificial media without producing fruit body.

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A Study on Stress Corrosion Cracking of Fiber Reinforced Composite by Slow Strain Rate Test (저변형률시험법에 의한 섬유강화 복합재료의 응력부식균열에 관한 연구)

  • Lim, Jae-Gyu;Choi, Tae-Su
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.20 no.11
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    • pp.3433-3440
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    • 1996
  • This paper was investigation of the stres corrosion cracking(SCC) mechanism and the properties of corrosion fracture surface of glass fiber reinforced plastics(GFRP) produced by hand lay up(HLU) method in synthetic sea water. Test material is GFRP, that was used vinylester type epoxy acrylate resin and an unsaturated polyester as the matrix and the chopped strand mat(CSM) type E-glss fiber as the reinforcement. The slow strain rate test(SSRT) was performed on dry, wet and saturated wet specimens in sea water. Here the pH concentration of synthetic sea water was 8.2 and the strain rate is 1 x $10^{-6}$($sec^{-1}$) and test temperature ranges varied from $-60^{\circ}C$ to $80^{\circ}C$. It could be confirmed the fact that wet specimens tested at a particular test temperature ranges were appeared the eviences of SCC such as con-planar, mirror and hackle zone. Moreover, SCC of GFRP in sea water was characterised by falt fracture surfaces with only small amounts of fiber pull-out, in partial.