• 한국미생물·생명공학회지
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• 제50권3호
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• pp.422-429
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• 2022

The hepatitis C virus (HCV) genome contains a positive-sense single-stranded RNA molecule, and it is classified into 8 genotypes and 87 subtypes. Globally, over 350,000 people die from liver cirrhosis and hepatocellular carcinoma caused by HCV each year. Here, the genotype distribution of HCV was estimated in the population in Cheonan, Korea using Sanger sequencing. In addition, the correlation between HCV RNA level and genotype was assessed using real-time polymerase chain reaction (PCR); similarly, the correlation of HCV RNA level with isolation year (2007-2016) was determined using 463 consecutive serum samples obtained from patients at Dankook University Hospital, Cheonan, Korea. In 2007, genotype 1b (54.2%) was predominant, followed by genotypes 2a (41.7%), 1a (2.1%) and 3a (2.1%); whereas in 2016, the predominant genotype was 2a (49.0%), followed by genotypes 1b (46.9%), 3b (2%), and 4a (2%). Neither age nor sex was correlated with HCV genotype. Furthermore, the mean HCV RNA level decreased significantly from 2012 to 2016 (p < 0.05). However, no significant correlations between genotype and HCV RNA level were found. Overall, the findings revealed that genotypes 2a and 1b were the most common in Cheonan, and the prevalence of HCV genotype 1b tended to decrease over the past decade.

• 한국식품위생안전성학회지
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• 제33권4호
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• pp.280-288
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• 2018

본 연구는 국내에서 생산되거나 해외에서 수입되어 국내에서 유통되는 수산물 중에서 두족류를 문어류, 낙지류, 오징어류, 주꾸미류, 꼴뚜기류의 5개 그룹으로 구분하여 분석하였다. 두족류 5개 그룹을 판별을 하기 위해 미토콘드리아에 존재하는 유전자를 분석하였고, 그 중에서 COI (mitochondrial cytochrome C oxidase subunit I), 16s rRNA (16s ribosomal RNA), 12s rRNA (12s ribosomal RNA) 내에서 상당히 유사한 DNA 서열 부분과 일부 서열 변화 부분이 확인되었다. 명확하게 두족류 5개 그룹 판별을 하기 위해 COI, 16s rRNA, 12s rRNA 유전자의 일부 서열 변화 부분에서 그룹 특이적 프라이머 세트를 디자인하였다. 국내 외에서 확보한 두족류 시료(참문어, 낙지, 살오징어, 아메리카 대왕오징어, 갑오징어, 주꾸미, 모래주꾸미, 하이야주꾸미, 참꼴뚜기, 창꼴뚜기, 한치꼴뚜기)의 genomic DNA을 추출하여 각 그룹의 특이적 프라이머를 이용하여 SYBR 기반의 real-time PCR 시스템에 의해 분석되었고, threshold cycle (Ct) value와 같은 real-time PCR 결과 분석에 의해 두족류 내 그룹 판별이 가능하였다(Table 3).

• The Plant Pathology Journal
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• 제35권2호
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• pp.164-171
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• 2019

An assay for detecting Apple scar skin viroid (ASSVd) was developed based on nucleic acid sequence based amplification (NASBA) in combination with realtime detection during the amplification process using molecular beacon. The ASSVd specific primers for amplification of the viroid RNA and molecular beacon for detecting the viroid were designed based on highly conserved regions of several ASSVd sequences including Korean isolate. The assay had a detection range of $1{\times}10^4$ to $1{\times}10^{12}$ ASSVd RNA $copies/{\mu}l$ with reproducibility and precision. Following the construction of standard curves based on time to positive (TTP) value for the serial dilutions ranging from $1{\times}10^7$ to $1{\times}10^{12}$ copies of the recombinant plasmid, a standard regression line was constructed by plotting the TTP values versus the logarithm of the starting ASSVd RNA copy number of 10-fold dilutions each. Compared to the established RT-PCR methods, our method was more sensitive for detecting ASSVd. The real-time quantitative NASBA method will be fast, sensitive, and reliable for routine diagnosis and selection of viroid-free stock materials. Furthermore, real-time quantitative NASBA may be especially useful for detecting low levels in apple trees with early viroid-infection stage and for monitoring the influence on tree growth.

• Asian Pacific Journal of Cancer Prevention
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• 제13권10호
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• pp.5149-5153
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• 2012

Introduction: This study assessed the relationship of E-cadherin mRNA and protein expression with the diagnosis of lung cancer with the aim of providing an auxiliary diagnostic method. Methods: Semi-quantitative nested RT-PCR and western blotting were applied to detect E-cadherin mRNA transcripts and protein, respectively, in 30 cases of diagnostic lung cancer, 30 cases of clinically suspected patients with lung cancer and 30 cases of other disease. Immunohistochemical staining was also used to detect E-cadherin. Results: Remarkably decreased levels of relative E-cadherin mRNA value and increased E-cadherin protein negativity were observed in probable lung cancer, when compared with possible lung cancer and others. With a threshold of 1.45, relative E-cadherin mRNA value showed a sensitivity of 90% and a specifity of 83% for the diagnosis of lung cancer. The combination of decreased relative E-cadherin mRNA value and negative E-cadherin protein increased the specificity and sensitivity. Conclusion: These data suggest that Chinese patients with diagnostic lung cancer have similar decreased levels of relative E-cadherin mRNA and E-cadherin protein value in the lung cancer tissues as in lung cancer patients in other countries. Measurement of relative E-cadherin mRNA and protein values in lung cancer tissues has potential for lung cancer diagnosis.

• 한국식품위생안전성학회지
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• 제33권1호
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• pp.71-76
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• 2018

노로바이러스는 전 세계적으로 산발적인 발병 관련 비세균성 위장염의 주요 원인 물질이다. 노로바이러스 검출을 위해 역전사 실시간 PCR (RT qPCR)이 그 민감도와 특이성으로 인해 주요 수단으로 빠르게 자리 잡았다. 그러나 RT qPCR 분석을 위해서는 정확한 바이러스 RNA 추출방법이 필수적이다. TRIzol 시약은 생물학적 물질로부터 RNA의 추출에 이용되고 따라서 노로바이러스 RNA 추출에도 널리 사용된다. 이 연구에서는 인체 노로바이러스 유전체 그룹 I (GI) 및 유전자 그룹 II (GII)와 생쥐 노로바이러스(GV) 중에서 GII로부터의 TRIzol 을 이용한 바이러스 RNA의 추출률이 바이러스 시료 용액의 pH에 의존했다는 내용이 다루어졌다. 실시간 PCR의 Ct값으로 비교한 RNA 추출 수율은 산성 영역보다 알칼리성 pH에서 높았다. 이 연구 결과로 부터 TRIzol을 이용하여 GII RNA를 추출하여 노로바이러스를 정량적으로 분석할 때 pH조건이 대단히 중요하다는 것을 알 수 있었다.

• 대한수의학회지
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• 제7권2호
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• pp.13-18
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• 1967

1. Within experimental chromatin, the total protein: DNA ratio did not vary in the same organs of control and irradiated rats. However, the amount of RNA and total protein associated with the DNA varied considerably among the different types of chromatin. In particular, the content of chromatin was the control tissue. RNA and total protein ratio of chromatins from brtain, liver, testis and spleen declined with experimental I organs. 2. There was the same quantitative relationship between the amount of RNA and the amount of histone-protein associated with DNA in chromatin. 3. RNA: DNA ratio of chromatin showed 1.5-2 times increas in the irradiated organs except brain. However, RNA: DNA ratio was decreased in chromatin by irradiation. 4. Histone-protein:residual protein ratio was greatly varied among the organs. However, the effect was not found by irradiation. 5. Priming activity of chromatin showed a higher value in testis and the activity was greater in organs with higher metabolic activity: 6. Inhibition of Actinomycin D is observable in chromatin from testis, liver, spleen and brain declined without relationship between irradiated and non-irradiated conditions. Ammonium sulfate showed increased priming activity by the electrostatic dissociation of DNA and histone in chromatin on the stimulation depending on property of chromatins. 7. It is suggested that the results support a proposal that testis and spleen of highly sensitive to irradiation should an increase in the priming activity whereas brain and liver of lower sensitivity decreased in the activity.

• Animal cells and systems
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• 제5권3호
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• pp.217-224
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• 2001

The present study examines the expression and regulation of gonadotropin-releasing hormone (GnRH) and its receptor (GnRH-R) mRNA levels during mouse ovarian development. A fully processed, mature GnRH mRNA together with intron-containing primary transcripts was expressed in the immature mouse ovary as determined by Northern blot analysis and reverse transcription-polymerase chain reaction (RT-PCR). The size of ovarian GnRH mRNA was similar to that of hypothalamus, but its amount was much lower than that in the hypothalamus. Quantitative RT-PCR procedure also revealed the expression of GnRH-R mRNA in the ovary, but the estimated amount was a thousand-fold lower than that in the pituitary gland. We also examined the regulation of ovarian GnRH and GnRH-R mRNA levels during the follicular development induced by pregnant mare's serum gonadotropin (PMSG) and/or human chorionic gonadotropin (hCG). Ovarian luteinizing hormone receptor (LH-R) mRNA was abruptly increased st 48 h after the PMSG administration and rapidly decreased to the basal level thereafter. Ovarian GnRH mRNA level was slightly decreased at 48 h after the PMSG administration, and then returned to the basal value. GnRH-R mRNA level began to increase at 24 h after the PMSG treatment, decreased below the uninduced basal level at 48 h, and gradually increased thereafter. HCG administration did not alter ovarian GnRH mRNA level, while it blocked the PMSG-induced increase in GnRH mRNA level. Taken together, the present study demonstrates that the expression of GnRH and GnRH-R mRNA are regulated by gonadotropin during follicular development, suggesting possible intragonadal paracrine roles of GnRH and GnRH-R in the mouse ovarian development.

• 한국발생생물학회지:발생과생식
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• 제14권4호
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• pp.261-268
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• 2010

As in the O. mykiss electrophoretic profiles of RNA, the signals of each RNA sample from 9 individual tissues such as liver, muscle, brain, heart, pituitary gland, kidney, intestine, spleen and gill similar to positive control were obtained. The tissue distributions of the complimentary DNA (cDNA) of O. mykiss four genes were analyzed using quantitative real-time PCR with primer sets for tissue expression analysis. In this rainbow trout species, author obtained bands of various sizes, ranged from 700 bp to 1,400 bp. A dissociation curve was made at the end of each run to make sure that there was no non-specific amplification. Supplementarily, the Ct of each DNA was compared. The Ct values of vinculin with rainbow trout tissues were determined in a manner similar to those for agouti-related protein (AgRP) and melanocortin receptors (MC4R I and MC4R II). Further, obtained Cts for standard curve of each DNA were affected by specific product (vinculin, AgRP and MC4R II genes). After several experiments with four individual genes of rainbow trout, author estimated a variation ratio of the mean Ct value of the DNA extracted using the comparative CTt method was 37.27, and the standard deviation was 5.33. The correlation coefficient between the Ct values and the concentration of cDNA was -0.98, -0.99, -0.91 and -0.86, respectively (vinculin, AgRP, MC4R I and MC4R II genes). Since this correlation showed high linearity, the straight line obtained was used as a standard for the O. mykiss tissues reared in aquarium. A PCR efficiency of 100% is ideally achieved when the slopes are close to the theoretical value of -3.31. According to quantification method, the results of quantification are strongly affected by the DNA fragmentation. The size of most DNA fragments obtained from various tissues of rainbow trout used in the experiment was approximately 100 bp. According to the four slopes, an efficiency of nearly 100% was estimated for four genes detection methods. Additionally, further analysis with more individuals and primers will be required to fully establish optimization in rainbow trout.

• 대한핵의학회지
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• 제1권2호
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• pp.27-34
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• 1967

근년(近年) 고등동물세포(高等動物細胞)에 있어서 유전자(遺傳子)의 본체(本體)인 DNA에서 RNA를 경과(經過)해서 특이적(特異的)인 단백질(蛋白質)의 생합성(生合成)에 도달(到達)하는 경로(經路)에 대(對)해서는 많은 연구(硏究)에 의해서 확립(確立)되어졌으나 그 조절기구(調節機構)에 대(對)해서는 불명(不明)한 점(點)이 많다. 개체(個體), 기관(器管), 세포내구조(細胞內構造) 급(及) DNA의 준위(準位)에서의 방사선(放射線)의 장해(障害)에 대(對)해서도 연구(硏究)되고 있으나 소위(所謂) 방사선감수성(放射線感受性) 급(及) 비감수성(非感受性)의 각장기(各臟器)에서 분리(分離)한 Chromatin (DNA-Histone-잔여단백(殘餘蛋白)의 고차구조결합체(高次構造結合體)에 대(對)한 DNA, RNA, 전단백질(全蛋白質)과 유전수식체(遺傳修飾體)라고 생각되는 Histon-단백(蛋白)의 화학조성(化學組成)을 검출(檢出)했으며 겸(兼)해서 chromatin의 생물활성(生物活性)인 RNA 합성능(合成能)(priming activity)에 대(對)한 방사선(放射線)의 영향(影響)을 조사(調査)하는데 의의(意義)가 있다. 전리방사선(電離放射線) 조사(照射)에 의해서 생체(生體)의 DNA의 합성조해(合成阻害)가 잘 알려진 사실(事實)이나 분화(分化)한 생체조직(生體組織)에서의 DNA의 합성(合成)보다도 일반대사(一般代謝)에 중요(重要)한 역할(役割)을 한다는 것도 생각된다. 세포(細胞)의 대사(代謝)는 내분비계등(內分泌系等)의 "Effector-DNA-RNA-단백합성(蛋白合成)이라는 정보유전기구(情報遺傳機構)에 의해서 제어(制禦)되어 있다. 이 연구(硏究)는 방사선생물학상(放射線生物學上) 중요(重要)한 것은 논할(論) 필요(必要)도 없으며 방사선동위원소표지화합물(放射線同位元素標識化合物)을 사용(使用)하여 생화학적(生化學的)으로 추구(推究)하였다.

• 한국발생생물학회지:발생과생식
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• 제27권2호
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• pp.91-99
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• 2023

The sea cucumber, Apostichopus japonicus, is one of the most valuable aquatic species. The color of body wall and appearance are important for the value of sea cucumbers. To examine expression pattern of long-chain acyl-coenzyme A dehydrogenase (LCAD), nuclear distribution C-containing protein 3 (NUDCD3), and receptor tyrosine kinase Tie-1 (TIE1), previously reported as differently expressed genes during the pigmentation of sea cucumber, we analyzed the temporal profiles of LCAD, NUDCD3, and TIE1 mRNAs in LED-exposed and light-shielded A. japonicus. Real-time quantitative PCR revealed that the LCAD, NUDCD3, and TIE1 mRNAs from the juveniles at 40-60 days post-fertilization (dpf) exhibited increasing patterns as compared to those of an early developmental larva (6-dpf). At 60-dpf juveniles, the LCAD and TIE1 mRNA levels of LED-exposed individuals were higher than those of light-shielded ones, whereas at 40-dpf and 50-dpf juveniles, the NUDCD3 mRNA expression was higher in the light-shielded condition (p<0.05). In the pigmented juveniles (90-dpf), the LCAD and TIE1 mRNA levels tended to show higher levels in red individuals than those in green ones, but there was a conversely higher level of NUDCD3 mRNA in green larva. In situ examination of LCAD and NUDCD3 mRNAs in light-shielded 6-dpf larva revealed that both genes are mainly expressed in the internal organs compared to the body surface. Together, these results may provide insights into the differential gene expression of LCAD, NUDCD3, and TIE1 during pigmentation process of the sea cucumber.