• Title/Summary/Keyword: Restriction sites

Search Result 183, Processing Time 0.035 seconds

Comparison of Glucuronidating Activity of Two Human cDNAs, UDPGTh1 and UDPGTh2

  • Kim, Soon-Sun;Owens, Ida-S.;Sheen, Yhun-Yhong
    • Archives of Pharmacal Research
    • /
    • v.20 no.5
    • /
    • pp.454-458
    • /
    • 1997
  • Two human liver UDP-glucuronosyltransferase cDNA clones, HLUG25 and UDPGTh2 were previously shown to encode isozymes active in the glucuronidation of hyodeoxycholic acid (HDCA) and certain estrogen derivatives (e.g., estriol and 3,4-catechol estrogens), respectively. in this study we have found that the UDPGTh2-encoded isoform (UDPGTh2) and HLUG25-encoded isoform (UDPGThl) have parallel aglycone specificities. When expressed in COS 1 cells, each isoform metabolized three types of dihydroxy- or trihydroxy-substituted ring structures, including the 3,4-catechol estrogen (4-hydroxyestrone), estriol, 17-epiestriol, and HDCA, but the UDPGTh2 isozyme was 100-fold more efficient than UDPGTh1. UDPGTh1 and UDPGTh2 were 86% identical overall (76 differences out of 528 amino acids), including 55 differences in the first 300 amino acids of the amino terminus, a domain which conferred the substrate specificity. The data indicated that a high level of conservation in the amino terminus was not required for the preservation of substrate selectivity. Analysis of glucuronidation activity encoded by UDPGTh1/UDPGTh2 chimeric cDNA constructed at their common restriction sites, Sac I (codon 297), Nco I (codon 385), and Hha I (codon 469), showed that nine amino acids between residues 385 and 469 were important for catalytic efficiency, suggesting that this region represented a domain which was critical for the catalysis but distinct from that responsible for aglycone-selection. These data indicate that UDPGTh2 is a primary isoform responsible for the detoxification of the bile salt intermediate as well as the active estrogen intermediates.

  • PDF

Associations of IL-10 Gene Polymorphisms with Acute Myeloid Leukemia in Hunan, China

  • Yao, Chen-Jiao;Du, Wei;Chen, Hai-Bing;Xiao, Sheng;Wang, Cheng-Hong;Fan, Zi-Li
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.4
    • /
    • pp.2439-2442
    • /
    • 2013
  • We investigated the possible association of interleukin-10 (IL-10) single nucleotide polymorphisms (SNPs) and susceptibility to acute myeloid leukemia (AML) in 115 patients and 137 healthy controls. Genetic analysis of IL-10 SNPs at -819 and -592 was carried out with the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) approach. The IL-10 mRNA expression of AML patients and controls with different genotype was detected by real-time quantitative polymerase chain reaction (RT-PCR). Genetic analysis of IL-10 revealed that the -819AA genotype frequencies and the -819A allele frequencies in the AML group were higher than in the controls (59.1% vs 40.9%; 75.6% vs 63.9%, respectively); there were remarkable differences in -819T/C and -592A/C gene distribution (P<0.05) and the TA haploid frequencies were higher in the AML group (75.6% vs 63.9%, P<0.05). IL-10 mRNA expression in incipient AML patients was obvious higher than the non-tumor group and the remission group ($7.78{\times}10^{-3}$ vs $2.43{\times}10^{-3}$, $3.64{\times}10^{-3}$, P<0.05).The study suggested that the haploid TA and genotype TA/TA may be associated with AML in Han people in Hunan province.The IL-10 SNPs at -819 and -592 sites were associated with AML and may affect IL-10 mRNA expression in AML patients.

Vitamin D Receptor Gene TaqI, BsmI and FokI Polymorphisms in Korean Patients with Tuberculosis

  • Kang, Tae-Jin;Jin, Song-Hou;Yeum, Chung-Eun;Lee, Seong-Beom;Kim, Chi-Hong;Lee, Sang-Haak;Kim, Kwan-Hyoung;Shin, Eun-Soon;Chae, Gue-Tae
    • IMMUNE NETWORK
    • /
    • v.11 no.5
    • /
    • pp.253-257
    • /
    • 2011
  • Background: The active metabolite (1, 25- dihydroxycholecalciferol) of vitamin D (25-hydroxycholecalciferol) leads to activation of macrophages and deficiency of vitamin D seems to be involved in the risk of tuberculosis. The effects of vitamin D are exerted by interaction with the vitamin D receptor (VDR) and may be influenced by polymorphism in the VDR gene. In this study, variation in the VDR gene was investigated in Korean population with tuberculosis. Methods: We typed three VDR polymorphisms of restriction endonuclease sites for TaqI, BsmI and FokI in 155 patients with tuberculosis and 105 healthy volunteers. Results: The frequencies of FokI genotypes determined from TB patients were 29.13% for FF, 56.31% for Ff, and 14.56% for ff. We observed 1.4-fold increased prevalence of the Ff genotype in TB patients compared with normal healthy groups (p=0.0857). However, there was no significant association between the genotype groups, TB patient and normal control, for FokI polymorphism. There was also no significant association between VDR gene and tuberculosis in another polymorphism (BsmI and TaqI). Conclusion: Three polymorphisms (TaqI, BsmI and FokI) in the VDR gene do not appear to be responsible for host susceptibility to human tuberculosis in Korean population.

Preparation and Antioxidant Activities In Vitro of a Designed Antioxidant Peptide from Pinctada fucata by Recombinant Escherichia coli

  • Wu, Yanyan;Ma, Yongkai;Li, Laihao;Yang, Xianqing
    • Journal of Microbiology and Biotechnology
    • /
    • v.28 no.1
    • /
    • pp.1-11
    • /
    • 2018
  • An antioxidant peptide derived from Pinctada fucata meat using an Alcalase2.4L enzymatic hydrolysis method (named AOP) and identified by LC-TOF-MS has promising clinical potential for generating cosmetic products that protect skin from sunshine. To date, there have been few published studies investigating the structure-activity relationship in these peptides. To prepare antioxidant peptides better and improve their stability, the design and expression of an antioxidant peptide from Pinctada fucata (named DSAOP) was studied. The peptide contains a common precursor of an expression vector containing an ${\alpha}$-helix tandemly linked according to the BamHI restriction sites. The DNA fragments encoding DSAOP were synthesized and subcloned into the expression vector pET-30a (+), and the peptide was expressed mostly as soluble protein in recombinant Escherichia coli. Meanwhile, the DPPH radical scavenging activity, superoxide radical scavenging activity, and hydroxyl radical scavenging activity of DSAOP $IC_{50}$ values were $0.136{\pm}0.006$, $0.625{\pm}0.025$, and $0.306{\pm}0.015mg/ml$, respectively, with 2-fold higher DPPH radical scavenging activity compared with chemosynthesized AOP (p < 0.05), as well as higher superoxide radical scavenging activity compared with natural AOP (p < 0.05). This preparation method was at the international advanced level. Furthermore, pilot-scale production results showed that DSAOP was expressed successfully in fermenter cultures, which indicated that the design strategy and expression methods would be useful for obtaining substantial amounts of stable peptides at low costs. These results showed that DSAOP produced with recombinant Escherichia coli could be useful in cosmetic skin care products, health foods, and pharmaceuticals.

Landscape Awareness Survey for Improvement of Urban Landscape in Yong-in city - Focusing on Residents, Government Officials, and Developers- (용인시 도시경관 개선을 위한 경관의식 조사 - 시민, 공무원, 개발사업자를 대상으로 -)

  • Sung, Hyun-Chan
    • Journal of the Korean Society of Environmental Restoration Technology
    • /
    • v.6 no.6
    • /
    • pp.1-13
    • /
    • 2003
  • This study aims at conducting an awareness survey of three parties, namely, citizens, government officials, and developers for improvement of urban landscape and, based on the survey result, to suggest a direction to develop a landscape plan for improvement of urban landscape of Yong-in city in the future. The summary of this study is as follows. First, in case of Yong-in, the main drivers of forest landscape destruction were "residential complexes developed for apartment buildings" and "golf courses", the driver of river landscape damage was "the status of cleanliness of riverside and poor facilities", the driver of roadside landscape damage was "construction sites" that completely eliminate original natural landscape in both citizen and government official groups. When granting development-related approvals in the future, appropriate actions should be prepared such as developing a landscape management guideline or including a landscape plan in advance to assure plans and designs fully reflecting landscape. Second, citizens' commitment to participate in urban landscape creation and improvement was rated high at even willing to tolerate regulations including restriction of property rights. In the future, landscape improvement programs allowing resident participation should be developed proactively. The roles of government officials include preparing various criteria at a city level and convincing and guiding citizens to initiate landscape improvement project together with citizens and developers through proactive efforts by government officials. Third, the role of developers would be to realize development projects conserving local environment and/or harmonizing with local natural environment before preparing a development plan or launching construction rather than offering various considerations after executing development projects.

Cloning and Expression of pcbAB Genes from Pseudomonas sp. DJ-12 in Escherichia coli (Pseudomonas sp. DJ-12 pcbAB 유전자의 Escherichia coli에서의 클로닝 및 발현)

  • 한재진;성태경;김치경
    • Korean Journal of Microbiology
    • /
    • v.31 no.2
    • /
    • pp.129-134
    • /
    • 1993
  • The pchAB genes of Pseudomonas sp. DJ-12 produce the enzymes of 4-chlorobipheny] (4CB) dioxygenase and dihydrodiol dehydrogenase which act on the first and second steps in degradation of 4CB and biphenyl. The genes were cloned in E coli XLI-Blue. The pcbAB genes of about 2.2 kb in size were contained in the pCUlO1 hybrid plasmid in the cloned cell of CUIOI. The genes were found to have their own promoter and three restriction sites for HindlII. 2,3-dihydroxybiphenyl was detected by the resting cell assay, as the metabolite transformed from biphenyl by the cloned cell of CUIOI. This means that the pcbAB genes are well expressed in E. coli. But dechlorination was unlikely involved in the pchAB gene expression but was believed to occur by functioning on 4CBA produced after ring-cleavage of 4CB.

  • PDF

Assessment of Applicability of Pretentioned Soil-Nail Systems with in-situ monitoring (현장 계측을 통한 프리텐션 쏘일네일링 시스템의 적용성 평가)

  • Lee, Hyuk-Jin;Ahn, Kwang-Kuk;Kim, Hong-Taek;Bang, Yoon-Kyung
    • Proceedings of the Korean Geotechical Society Conference
    • /
    • 2005.10a
    • /
    • pp.320-329
    • /
    • 2005
  • The use of diverse methods for the retaining system has been continuously increased in order to maintain the stability during excavation. However, ground anchor system occasionally may have the restriction in urban excavation sites nearby the existing structures because of space limitation. In this case, soil nailing system with relatively short length of nails could be efficiently useful as an alternative method. The general soil nailing support system, however, may result in excessive deformations particularly in excavating the zone of weak soils or nearby the existing structures. Therefore, applying the pretension force to the soil nails then could play important roles to reduce deformations mainly in an upper part of the nailed-soil excavation system as well as to improve the local slope stability. In this study, a newly modified soil nailing technology named as the PSN(Pretention Soil Nailing) is developed to reduce both facing displacements and ground surface settlements during top-down excavation process as well as to increase the global slope stability. Up to now, the PSN system has been investigated mainly focusing on an establishment of the design procedure. In the present study, the field tests including pull-out tests were fulfilled to investigate the behavior of characteristics for PSN system. All results of tests were also analyzed to provide a fundamental and efficient design.

  • PDF

Tobamoviruses of Green Peppers Growing on Hydroponic Systems (풋고추 수경재배에서 발생하는 tobamovirus의 특성)

  • Choi, Gug-Seoun;Kim, Jae-Hyun;Kim, Jeong-Soo;Kim, Hyun-Ran
    • Research in Plant Disease
    • /
    • v.10 no.3
    • /
    • pp.194-197
    • /
    • 2004
  • Incidence of tobamovirus diseases was 100% at late growth stage of green pepper on hydroponic systems in plastic house. Infection frequency of the diseases showed 34% of Pepper mild mottle virus (PMMoV), 41.5% of Tobacco mild green mottle virus (TMGMV), and 24.5% of the co-infected viruses. The two viruses specifically reacted in DAS-ELISA prepared with each polyclonal antibody. A total of 77 pure tobamovirus isolates obtained from the crops was tested for pathotype determination. The isolation frequency of tobamovirus pathotype $P_{0}$ and $P_{1,2}$ was 61 % and 39%, respectively. All TMGMV isolates belonged to the pathotype $P_{0}$. In restriction enzyme analysis of the cDNAs synthesized with coat protein gene of PMMoV pathotype $P_{0}$ and $P_{1,2}$, the former had two TaqI sites but the later had one.one.

A study on the stability analysis for asymmetry parallel tunnel with rock pillar (암반 필라를 포함한 비대칭 근접 병설터널의 안정성 평가에 관한 연구)

  • Kim, Do-Sik;Kim, Young-Geun
    • Journal of Korean Tunnelling and Underground Space Association
    • /
    • v.9 no.4
    • /
    • pp.387-401
    • /
    • 2007
  • Recently, because of the restriction of land for construction and interference of adjacent structure, parallel tunnels with small clearance have been planned and constructed in many sites. In this case, the stability of pillar at center part is very important factor to satisfy the stability of tunnel structure under the construction. In this paper, numerical analyses for the asymmetry parallel tunnels with a narrow width of pillar have been carried out to search for the optimum reinforcement measure for rock pillar and verify the stability of tunnel. Rock pillar between each single tunnel is supposed to be under heavy load by rock mass. The analysis of stress state at rock pillar at various cases for construction conditions is required to investigate the structural behaviour of tunnels and stability of the pillar. Strength-stress ratio is calculated based on the failure theory of rock and the safety factor of tunnel is computed with strength reduction technique. Through these numerical results, reasonable reinforcement measures for rock pillar at parallel tunnel were established and recommended.

  • PDF

Alu Methylation in Serum from Patients with Nasopharyngeal Carcinoma

  • Tiwawech, Danai;Srisuttee, Ratakorn;Rattanatanyong, Prakasit;Puttipanyalears, Charoenchai;Kitkumthorn, Nakarin;Mutirangura, Apiwat
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.15 no.22
    • /
    • pp.9797-9800
    • /
    • 2014
  • Background: Nasopharyngeal carcinoma (NPC) is a common cancer in Southern China and Southeast Asia. Alu elements are among the most prevalent repetitive sequences and constitute 11% of the human genome. Although Alu methylation has been evaluated in many types of cancer, few studies have examined the levels of this modification in serum from NPC patients. Objective: To compare the Alu methylation levels and patterns between serum from NPC patients and normal controls. Materials and Methods: Sera from 50 NPC patients and 140 controls were examined. Quantitative combined bisulfite restriction analysis-Alu (qCOBRA-Alu) was applied to measure Alu methylation levels and characterize Alu methylation patterns. Amplified products were classified into four patterns according to the methylation status of 2 CpG sites: hypermethylated (methylation at both loci), partially methylated (methylation of either of the two loci), and hypomethylated (unmethylated at both loci). Results: A comparison of normal control sera with NPC sera revealed that the latter presented a significantly lower methylation level (p=0.0002) and a significantly higher percentage of hypomethylated loci (p=0.0002). The sensitivity of the higher percentage of Alu hypomethyted loci for distinguishing NPC patients from normal controls was 96%. Conclusions: Alu elements in the circulating DNA of NPC patients are hypomethylated. Moreover, Alu hypomethylated loci may represent a potential biomarker for NPC screening.