• Title/Summary/Keyword: circular RNA

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Complete genome sequence of probiotic Lactobacillus johnsonii 7409N31 isolated from a healthy Hanwoo calf

  • Young Joon Oh;Jieun Lee;Seul Ki Lim;Min-Sung Kwon;Sulhee Lee;Sang-Pil Choi;Dohyeon Yu;Yeon-su Oh;Jinho Park;Hak-Jong Choi
    • Journal of Animal Science and Technology
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    • v.65 no.4
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    • pp.890-893
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    • 2023
  • Lactobacillus johnsonii 7409N31 was isolated from the feces of a healthy 11-day-old Hanwoo calf from a farm in Geochang-gun, Gyeongsangnam-do, Korea. The genome of the strain was completely sequenced using the PacBio RSII sequencing system, and it was confirmed that it was composed of one circular chromosome. The size of the entire genome was 2,198,442 bp, and it had 35.01 mol% guanine + cytosine (G + C) content and 2,222 protein-coding sequences, 24 rRNA, 3 ncRNA, and 112 tRNA genes. Strain 7409N31 possessed genes encoding enzymes involved in the hydrolysis of both fibrous and non-fibrous carbohydrates. These data provide a comprehensive theoretical understanding for developing industrial probiotic feed additives that improve nutrient digestibility.

Occurrence of Apple scar viroid-Korean strain (ASSVd-K) in Apples Cultivated in Korea

  • Lee, Ju-Hee;Park, Jean-Kyung;Lee, Dong-Hyuk;Uhm, Jae-Youl;Ghim, Sa-Youl;Lee, Jai-Youl
    • The Plant Pathology Journal
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    • v.17 no.5
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    • pp.300-304
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    • 2001
  • Apple is the most economically important fruit in Korea. The suspected viroid disease of dapple apple was found in apple fruits cultivated in Kyungpook province. Symptoms begin in mid-July as small circular spots, which stand out against the background color on the young fruit. Dappling of the fruit becomes more intense and easier to detect as the fruit approaches maturity; the affected spots remain yellowish as the fruit matures. no leaf or bark syndromes have been associated with this disease. The infected fruits are downgraded considerably during quality grading. The low molecular weight RNA containing viroid RNA molecules were extracted from the peels of the apples with dapple symptoms. The RNA molecules were extracted from the apples using Qiagen column chromatography. The purified RNAs were used for the synthesis of cDNA with RT-PCR. The PCR products were then ligated into a pGEM-T Easy vector, cloned and sequenced. The sequence of the viroid RNA molecule shows 331 nucleotides with one base difference ("G" insertion between the position of 133 and 134) compared with that of the Apple scar skin viroid (ASSVd) reported by Hashimoto and Koganezawa in Japan. This is the first report on the occurrence of the ASSVd in apple trees cultivated in Korea, as well as the identification of a new Korean strain of the ASSVd.the ASSVd.

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Synthesis and base pairing properties of DNA-RNA heteroduplex containing 5-hydroxyuridine

  • Cui, Song;Kim, Yong-Hoon;Jin, Cheng-Hao;Kim, Sang-Kook;Rhee, Man-hee;Kwon, Oh-Shin;Moon, Byung-Jo
    • BMB Reports
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    • v.42 no.6
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    • pp.373-379
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    • 2009
  • 5-Hydroxyuridine (5-OHU) is a major lesion of uridine and cytosine produced in RNA by various chemical oxidants. To elucidate its biochemical and biophysical effects on RNA replication, the site-specifically modified oligoribonucleotides containing 5-OHU were synthesized with C5-hydroxy-5'-ODMTr-2'-TBDMS-uridine phosphoramidite using automated solid phase synthesis. The base-pairing properties of nucleotides opposite 5-OHU in 24 mer oligoribonulcleotides with dNTP were studied using three reverse transcriptases (Super-$Script^{TM}II$-, AMV-, MMLV-RT) in cDNA synthesis. Adenine as well as guanine was incorporated preferentially by all reverse transcriptases. In the UV-melting temperature experiment, the results from the relative stabilities of the base pairs were A : 5-OHU > G : 5-OHU > T : 5-OHU $\approx$ C : 5-OHU. Circular Dichroism (CD) studies showed that DNA-RNA containing 5- OHU heteroduplexes exhibit a similar conformation between the A-type RNA and B-type DNA. These results suggest that 5- OHU from oxidative damage was mainly influenced by adenine mismatch.

CircCOL1A2 Sponges MiR-1286 to Promote Cell Invasion and Migration of Gastric Cancer by Elevating Expression of USP10 to Downregulate RFC2 Ubiquitination Level

  • Li, Hang;Chai, Lixin;Ding, Zujun;He, Huabo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.7
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    • pp.938-948
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    • 2022
  • Gastric cancers (GC) are generally malignant tumors, occurring with high incidence and threatening public health around the world. Circular RNAs (circRNAs) play crucial roles in modulating various cancers, including GC. However, the functions of circRNAs and their regulatory mechanism in colorectal cancer (CRC) remain largely unknown. This study focuses on both the role of circCOL1A2 in CRC progression as well as its downstream molecular mechanism. Quantitative polymerase chain reaction (qPCR) and western blot were adopted for gene expression analysis. Functional experiments were performed to study the biological functions. Fluorescence in situ hybridization (FISH) and subcellular fraction assays were employed to detect the subcellular distribution. Luciferase reporter, RNA-binding protein immunoprecipitation (RIP), co-immunoprecipitation (Co-IP), RNA pull-down, and immunofluorescence (IF) and immunoprecipitation (IP) assays were used to explore the underlying mechanisms. Our results found circCOL1A2 to be not only upregulated in GC cells, but that it also propels the migration and invasion of GC cells. CircCOL1A2 functions as a competing endogenous RNA (ceRNA) by sequestering microRNA-1286 (miR-1286) to modulate ubiquitin-specific peptidase 10 (USP10), which in turn spurs the migration and invasion of GC cells by regulating RFC2. In sum, CircCOL1A2 sponges miR-1286 to promote cell invasion and migration of GC by elevating the expression of USP10 to downregulate the level of RFC2 ubiquitination. Our study offers a potential novel target for the early diagnosis and treatment of GC.

Characterization of the first mitogenomes of the smallest fish in the world, Paedocypris progenetica, from peat swamp of Peninsular Malaysia, Selangor, and Perak

  • Hussin, NorJasmin;Azmir, Izzati Adilah;Esa, Yuzine;Ahmad, Amirrudin;Salleh, Faezah Mohd;Jahari, Puteri Nur Syahzanani;Munian, Kaviarasu;Gan, Han Ming
    • Genomics & Informatics
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    • v.20 no.1
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    • pp.12.1-12.7
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    • 2022
  • The two complete mitochondrial genomes (mitogenomes) of Paedocypris progenetica, the smallest fish in the world which belonged to the Cyprinidae family, were sequenced and assembled. The circular DNA molecules of mitogenomes P1-P. progenetica and S3-P. progenetica were 16,827 and 16,616 bp in length, respectively, and encoded 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and one control region. The gene arrangements of P. progenetica were identical to those of other Paedocypris species. BLAST and phylogenetic analyses revealed variations in the mitogenome sequences of two Paedocypris species from Perak and Selangor. The circular DNA molecule of P. progenetica yield a standard vertebrate gene arrangement and an overall nucleotide composition of A 33.0%, T 27.2%, C 23.5%, and G 15.5%. The overall AT content of this species was consistent with that of other species in other genera. The negative GC-skew and positive AT-skew of the control region in P. progenetica indicated rich genetic variability and AT nucleotide bias, respectively. The results of this study provide genomic variation information and enhance the understanding of the mitogenome of P. progenetica. They could later deliver highly valuable new insight into data for phylogenetic analysis and population genetics.

RNA Interference of Chitinase Gene in Spodoptera litura (담배거세미나방(Spodoptera litura) Chitinase gene의 RNA interference)

  • Jeon, Mi Jin;Seo, Mi Ja;Youn, Young Nam;Yu, Yong Man
    • The Korean Journal of Pesticide Science
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    • v.18 no.3
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    • pp.202-209
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    • 2014
  • RNA interference (RNAi) is the method which controls phenotypes of gene in live cells. Chitinase is the enzyme helping digestion and absorption of old cuticles during the ecdysis of insects. In order to investigate molting-inhibition effect with the chitinase related gene in Spodoptera litura, RNA was extracted from the $5^{th}$ instars. cDNA was synthesized and then we obtained about 700 bp size chitinase. After PCR products were cloned into a pGEM T-easy vector, colonies were picked. DNA was extracted from the colony cultures. EcoR I enzyme was used to check whether PCR products were inserted or not. And then we confirmed vector band of about 3 kb and insert band of about 700 bp. To synthesize the dsRNA, each DNA was cut with Spe I and Nco I enzymes (Circular DNA became lineared DNA). After synthesis of dsRNA, approximately 5 ul dsRNA was injected into the $3^{rd}$ abdominal segment of S. litura $4^{th}$ larvae. The concentration of dsRNA was about $10{\mu}g/{\mu}l$. We confirmed larval-larval molting : there were phenotypically abnormal individuals - for instance malformation, molting inhibition and change of integument color. Pupaadult molting : there were phenotypically abnormal individuals - for instance molting inhibition, change of wings and malformation. Also we could investigate the pupation, emergence and variation about noninjection, treated with DW and dsRNA. Each pupation was non-injection 83.3%, DW 78.3% and dsRNA 66.7%. Each emergence was non-injection 90.0%, DW 72.3% and dsRNA 65.0%. So we considered that chitinase dsRNA induced molting inhibition effect. But each variation was non-injection 8.9%, DW 2.9% and dsRNA 19.2%. Therefore dsRNA group showed the highest variation value. When 18 hours after injecting dsRNA, we could obtain abnormal individual.

Draft genome sequences of Vibrio splendidus KCTC 11899BP, which produces hyaluronate lyase in the presence of hyaluronic acid (히알우론산 유도하에 히알우로네이트 라이아제를 생산하는 Vibrio splendidus KCTC 11899BP균주의 유전체 서열 분석)

  • Park, Joo Woong;Lee, Sang-Eun;Shin, Woon-Seob;Kim, Kyoung Jin;Kim, Youn Uck
    • Korean Journal of Microbiology
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    • v.54 no.3
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    • pp.302-304
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    • 2018
  • We, for the first time, isolated and identified a Vibrio splendidus KCTC 11899BP producing hyaluronate lyase from seawater. This enzyme is produced only when hyaluronic acid (HA) is added to the basal medium. Hyaluronate lyases are produced by microorganisms, which degrade the ${\beta}$-(1, 4) bond of HA to produce disaccharide. The genome of KCTC 11899BP, which consist of two circular contigs that are 3,522 kb (contig 1) long and 1,986 kb (contig 2) long respectively, as like other Vibrio sp. that contained 2 chromosomes. The genome included 4,700 predicted open reading frames, G + C content 44.12%, 137 tRNA genes, and 46 rRNA genes.

Complete genome sequence of Bacillus velezensis YC7010, an endophytic bacterium with plant growth promoting, antimicrobial and systemic resistance inducing activities in rice (식물생육촉진, 항균 및 저항성 유도 효과를 나타내는 내생세균 Bacillus velezensis YC7010의 유전체 염기서열)

  • Harun-Or-Rashid, Md.;Hwang, Jeong Hyeon;Chung, Young Ryun
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.329-331
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    • 2017
  • Bacillus velezensis YC7010 is an endophytic bacterium isolated from the rice rhizosphere in Jinju, Republic of Korea, with properties conductive to growth promotion, antibiosis and induced systemic resistance to significant, soil-borne rice fungal and bacterial pathogens. The genome of B. velezensis YC7010 comprises a 3,975,683 bp circular chromosome which consists of 3,790 protein-coding genes (86tRNA and 27rRNA genes). Based on genomic analysis, we identified genes involved in colonization and establishment inside the plant, biosynthesis of antibiotic compounds such as surfactin, plipapastatin, bacillibactin, and bacillaene, as well as the production of the phytohormones and volatile compounds which serve to promote the plants growth and development.

Evaluating the Efficacy of a Formalin Alternative in Fixing Pathological Tissues for Histological and Molecular Diagnoses

  • Min-A Je;Haneul Lee;Heechul Park;Dong Hyeok Kim;Yeongdon Ju;Jaewon Lim;Sunghyun Kim;Jungho Kim
    • Biomedical Science Letters
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    • v.29 no.1
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    • pp.48-52
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    • 2023
  • Formaldehyde use is associated with serious health risks, which can affect medical personnel and technicians. Therefore, we investigated the efficacy of an alternative fixative, with respect to two types of formalin fixatives, by hematoxylin and eosin (H&E) staining, periodic acid Schiff (PAS) staining, immunohistochemical (IHC) staining, and RNA extraction. For H&E staining, the circular nucleus was stained dark blue by the basic dye hematoxylin and the cytoplasm was stained red by the acid dye eosin in all three fixative samples. No difference was found in the Duksan General Science (DGS), Sigma-Aldrich, and Core-Fix fixative samples (Corebiotech) used to fix kidney tissue, after PAS staining. IHC staining showed that CD4 was significantly increased in the lippolysaccharide (LPS)-treated group compared to the control group (vehicle), confirming the changes in specific molecules. The quantity and quality of RNA from tissues fixed in the three types of fixatives were evaluated. The average concentration of RNA was 106 ng/µL and average purity at A 260/280 ratio was 1.7~2.0, regardless of fixative used. For quality of protein, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein was confirmed by Western blotting. In conclusion, Core-Fix can be used as a fixative for pathological tissues, in histological and molecular diagnoses.

Construction of Infectious cDNA Clone of a Chrysanthemum stunt viroid Korean Isolate

  • Yoon, Ju-Yeon;Cho, In-Sook;Choi, Gug-Seoun;Choi, Seung-Kook
    • The Plant Pathology Journal
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    • v.30 no.1
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    • pp.68-74
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    • 2014
  • Chrysanthemum stunt viroid (CSVd), a noncoding infectious RNA molecule, causes seriously economic losses of chrysanthemum for 3 or 4 years after its first infection. Monomeric cDNA clones of CSVd isolate SK1 (CSVd-SK1) were constructed in the plasmids pGEM-T easy vector and pUC19 vector. Linear positive-sense transcripts synthesized in vitro from the full-length monomeric cDNA clones of CSVd-SK1 could infect systemically tomato seedlings and chrysanthemum plants, suggesting that the linear CSVd RNA transcribed from the cDNA clones could be replicated as efficiently as circular CSVd in host species. However, direct inoculation of plasmid cDNA clones containing full-length monomeric cDNA of CSVd-SK1 failed to infect tomato and chrysanthemum and linear negative-sense transcripts from the plasmid DNAs were not infectious in the two plant species. The cDNA sequences of progeny viroid in systemically infected tomato and chrysanthemum showed a few substitutions at a specific nucleotide position, but there were no deletions and insertions in the sequences of the CSVd progeny from tomato and chrysanthemum plants.