• Title/Summary/Keyword: t-RFLP

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Rice genotype, parental lineage and physiological tolerance to soil salinity shapes the community structure of rice seed bacterial endophytes

  • Walitang, Denver I.;Kim, Kiyoon;Chatterjee, Poulami;Kang, Yeongyeong;Sa, Tongmin
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.342-342
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    • 2017
  • Rice seeds are a home to endophytic bacterial communities which serve as a source of the plant's endophytes. As rice undergo physiological and adaptive modifications through cross breeding in the process of attaining salinity tolerance, this may also lead to changes in the endophytic bacterial community especially those residing in the seeds. This study explores the community structure of seed bacterial endophytes as influenced by rice parental lineage, genotype and physiological adaptation to salinity stress. Endophytic bacterial diversity was studied through culture dependent technique, cloning and Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis. Results revealed considerably diverse communities of bacterial endophytes in the interior of rice seeds. The richness of ribotypes ranges from 5-14 T-RFs corresponding to major groups of bacterial endophytes in the seeds. Endophytic bacterial diversity of the salt-sensitive IR29 is significantly more diverse compared to those of salt-tolerant cultivars. Proteobacteria followed by Actinobacteria and Firmicutes dominated the overall endophytic bacterial communities of the indica rice seeds based on 16S rDNA analysis of clones and isolates. Community profiles show common ribotypes found in all cultivars of the indica subspecies representing potential core microbiota belonging to Curtobacterium, Flavobacterium, Enterobacter, Xanthomonas, Herbaspirillum, Microbacterium and Stenotrophomonas. Multivariate analysis showed that the bacterial endophytic community and diversity of rice seeds are mainly influenced by their host's genotype, physiological adaptation to salt stress and parental lineage.

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Association of the CYP17-34T/C Polymorphism with Pancreatic Cancer Risk

  • Hussain, Shahid;Bano, Raisa;Khan, Muhammad Tahir;Khan, Mohammad Haroon
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.sup3
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    • pp.71-75
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    • 2016
  • Pancreatic cancer is a leading cause of fatality worldwide. Several population studies have been conducted on genetic diagnosis of pancreatic cancer but the results from epidemiologic studies are very limited. CYP17A gene has a role in disease formation but its influence on pancreatic cancer is unclear. A polymorphism in the 5'UTR promoter region of CYP17A1-34T/C (A1/A2) has been associated with multiple cancers. The aim of the current study was to assess associations of this polymorphism and socio-demographic risk factors with pancreatic cancer. A total of 255 and 320 controls were enrolled in the study, and were genetically analyzed through PCR-RFLP. Statistical analysis was conducted with observed genotype frequencies and odds ratios (ORs) and 95% CIs were estimated using unconditional logistic regression. The impact of socio-demographic factors was accessed through Kaplen-Meir analysis. According to our results, the A2/A2 genotype was significantly associated with pancreatic cancer (OR=2.1, 95%CI = 1.3-3.5). Gender female (OR=2.6, 95%CI=1.8-3.7), age group 80s/80+ years (OR=2.2, 95% CI=1.2-4), smoking both former (OR=4.6, 95% CIs=2.5-8.8) and current (OR=3.6, 95% CI=2-6.7), and family history (OR=7.1; 95%CI = 4.6-11.4) were also found associated with increased risk. Current study suggests that along with established risk factors for pancreatic cancer CYP17A1-34T/C may play a role. However, on the basis of small sample size the argument cannot be fully endorsed and larger scale studies are recommended.

Current status on the molecular biological research for the origin of cultivated sweetpotato [Ipomoea batatas L. (Lam)] (재배종 고구마[Ipomoea batatas L. (Lam)]의 기원종에 관한 분자생물학적 연구 동향)

  • Lee, Shin Woo;Kim, Yun-Hee
    • Journal of Plant Biotechnology
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    • v.48 no.4
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    • pp.223-227
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    • 2021
  • Several hypotheses for the origin of cultivated sweetpotato [Ipomoea batatas L. (Lam)] have been suggested but the exact progenitor is still unknown. Based on the results of RFLP patterns, microsatellite markers, SNP markers, FISH analyses, and genome analyses of haplotypes, wild species belonging to batatas group, I. trifida, I. leucantha, I. littoralis, I. tabascana, I. tenuissima, I. tiliacea, and I. triloba have been suggested as a progenitor. However, recently, advanced genomic technologies and characterization of the inserted T-DNA fragments of Agrobacterium in the genome of cultivated sweetpotato and wild species through horizontal gene transfer suggest that there may be an older progenitor than the wild species suggested so far.

Bacterial Community Dynamics during Composting of Food Wastes (음식물 쓰레기 퇴비화 과정에 따른 세균군집 구조의 변화)

  • Shin, Ji-Hye;Lee, Jin-Woo;Nam, Ji-Hyun;Park, Se-Yong;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.148-154
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    • 2009
  • Composting is a biological process converting solid organic waste into valuable materials such as fertilizer. The change of bacterial populations in a composting reactor of food waste was investigated for 2 months. Based on shifts in temperature profile, the composting process could be divided into the first phase ($2^{\circ}C\sim55^{\circ}C$), the second phase ($55^{\circ}C\sim97^{\circ}C$), and the third phase ($50^{\circ}C\sim89^{\circ}C$). The number of total bacteria was $1.66\times10^{11}$ cell/g, $0.29\times10^{11}$ cell/g, and $0.28\times10^{11}$ cell/g in the first, second, and third stages, respectively. The proportions of thermophiles increased from 33% to 89% in the second stage. T-RFLP analysis and nucleotide sequencing of 16S rRNA gene demonstrated that the change of bacterial community structure was coupled with shifts in composting stages. The structure of bacterial community in the ultra-thermophilic second stage reflected that of seeding starter. The major decomposers driving the ultra-thermophilic composting were identified as phylotypes related to Bacillus and Pseudomonas.

Isolation of dhlA Gene Responsible for Degradation of 1, 2-dichloroethane from Metagenomic Library Derived from Daecheong Reservoir (대청호로부터 제작한 메타지놈 라이브러리에서 1, 2-dichloroethane의 분해에 관여하는 dhlA 유전자의 분리)

  • Kang, Cheol-Hee;Moon, Mi-Sook;Song, Ji-Sook;Lee, Sang-Mhan;Kim, Chi-Kyung
    • Korean Journal of Ecology and Environment
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    • v.38 no.2 s.112
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    • pp.137-145
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    • 2005
  • Traditional screening techniques have missed up to 99% of microbial resources existing in the nature. Strategies of direct cloning of environmental DNAs comprising tine genetic blueprints of entire microbial metagenomes provide vastly more genetic information than is contained in the culturable. Therefore, one way to screening the useful gene in a variety of environments is the construction of metagenomic DNA library. In this study, the water samples were collected from Daecheong Reservoir in the mid Korea, and analyzed by T-RFLP to examine the diversity of the microbial communities. The crude DNAs were extracted by SDS-based freezing-thawing method and then further purified using an $UltraClean^{TM}kit$ (MoBio, USA). The metagenomic libraries were constructed with the DNAs partially digested with EcoRI, BamHI, and SacII in Escherichia coli DH10B using the pBACe3.6 vector. About 14.0 Mb of metagenomic libraries were obtained with average inserts 13 ${\sim}$ 15 kb in size. The genes responsible for degradation of 1, 2-dichloroethane (1, 2-DCE) via hydrolytic dehalogenation were identified from the metagenomic libraries by colony hybridization. The 1, 2-dichloroethane dehalogenase gene (dhlA) was cloned and its nucleotide sequence was analyzed. The activity of the 1, 2-DCE dehalogenase was highly expressed to the substrate. These results indicated that the dhlA gene identified from the metagenomes derived from Deacheong Reservoir might be useful to develop a potent strain for degradation of 1, 2-DCE.

Bacterial Community Structure of Food Wastewater Treatment System Combined with Rotating Biological Contactor and Tapered Aeration Reactor (회전접촉장치와 점감포기 반응조를 이용한 식품폐수 처리시설의 세균군집 구조)

  • Jeong, Soon-Jae;Nam, Ji-Hyun;Bae, Woo-Keun;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.169-176
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    • 2010
  • A pilot-scale wastewater treatment plant combined with rotating biological contactor and tapered aeration reactors was operated with the wastewater discharged from a food factory for 5 months. The bacterial communities of this plant were investigated by terminal restriction fragment length polymorphism (T-RFLP) and phylogenetic analysis of 16S rRNA genes. In spite of high concentration of nitrogen and phosphorus as well as organic carbon, removal efficiency of chemical oxygen demand, total nitrogen, and total phosphorus was 98%, 93%, and 95%, respectively. Bacterial community at the initial operation stage was clearly distinguished from that of the stable operation stage. The most predominant phylum in the sample of stable stage was Bacteroidetes. Major population of operation period was Haliscomenobacter, Sphaerotilus, and candidate division TM7, which were classified as filamentous bacteria. However, sludge bulking caused by these bacteria was not observed. The population that has a close relationship with Haliscomenobacter increased during the stable operation stage, emerging as the most predominant group. These results suggest that the filamentous bacteria participated in nutrient removal when using rotating biological contactor and tapered aeration reactor.

Nucleotide Sequencing and PCR-RFLP of Insulin-like Growth Factor Binding Protein-3 Gene in Riverine Buffalo (Bubalus bubalis)

  • Padma, B.;Kumar, Pushpendra;Choudhary, V.;Dhara, S.K.;Mishra, A.;Bhattacharya, T.K.;Bhushan, B.;Sharma, Arjava
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.7
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    • pp.910-913
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    • 2004
  • Insulin-like growth factor binding protein-3 (IGFBP-3) gene is a structural gene associated with the growth and development of the animals. The present investigation was carried out to unravel nucleotide sequence and polymerase chain reactionrestriction fragment polymorphism (PCR-RFLP) of IGFBP-3 gene in buffalo. Genomic DNA was isolated from a total of 157 animals belonging to Murrah, Surti, Jaffarabadi and Nagpuri breeds of Indian riverine buffalo. A 655 bp of IGFBP-3 gene was amplified in all the breeds and amplicons were digested with Hae III, Taq I and Msp I restriction enzymes. On digestion with Hae III yielded single restriction pattern of 8 fragments of sizes 201, 165, 154, 56, 36, 19, 16 and 8 bp in all the animals studied. Similarly Taq I and Msp I also revealed single restriction pattern yielding fragments of sizes 240 and 415 bp and 145 and 510 bp, respectively. This shows nonpolymorphic nature of restriction sites in buffalo. Nucleotide sequencing of 587 bp of IGFBP-3 gene in Murrah buffalo was done and submitted to the GenBank (Accession No. AY304829). Nucleotide sequencing revealed an addition of 4 bases in the intronic region as compared to cattle.

Analyses of Single Nucleotide Polymorphisms and Haplotype Linkage of the Human ABCB1 (MDR1) Gene in Korean

  • Ryu, Ho-Cheol;Kwon, Hyog-Young;Choi, Il-Kuen;Rhee, Dong-Kwon
    • Archives of Pharmacal Research
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    • v.29 no.12
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    • pp.1132-1139
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    • 2006
  • Single nucleotide polymorph isms (SNPs) in the MDR1 gene that are responsible for drug efflux can cause toxicity. Therefore, this study determined the SNPs of the Korean MDR1 gene, and analyzed the haplotypes and a linkage disequilibrium (LD) of the SNPs determined. The frequency of 9 SNPs from the MDR1 gene was determined by PCR-RFLP analyses of 100 to 500 healthy individuals. The frequcies of the SNPs were C3435T (47.7%), G2677T (37.6%), G2677A (4.4%), T1236C (21.7%), T129C (8%), A2956G (2.5%), T307C (1.5%), A41aG (9.2%), C145G (0%), and G4030C (0%). Analyses of the haplotype structure and an estimation of the LD of the combined polymorph isms demonstrated that the frequency of the 1236T-2677G-3435T haplotype is much higher in Koreans (14.1%) than in Chinese and western black Africans and the C3435T SNP in Koreans appears to have LD with T129C in Koreans for the first time. These results provide insight into the genetic variation of MDR1 in Koreans, and demonstrated the possibility of a new LD in this gene.