• 제목/요약/키워드: total RNA

검색결과 1,770건 처리시간 0.022초

저온순화 및 탈순화가 배추의 내동성 및 total RNA, soluble protein, soluble sugar 함량에 미치는 영향 (Effect of Cold Acclimation and Deacclimation on the Freezing Tolerance, Total RNA, Soluble Protein and Soluble Sugar in Chinese Cabbage)

  • Jeong Hyun Nam;Won Hee Kang;Il Seop Kim
    • 생물환경조절학회지
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    • 제10권4호
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    • pp.244-250
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    • 2001
  • 저온은 배추 묘에서 1주일동안 RNA함량과 단백질 함량을 증가시켰고, 당 함량에 있어서도 많은 변화를 유발했을 뿐만 아니라 저온순화시 치사온도에서 생존율 또한 증가하였다. 이런 결과에서 보듯이 이러한 변화는 내동성의 발달과 관련되어 있는 것으로 생각할 수 있다. 5$^{\circ}C$에서 처음 24시간처리 했을 때 12시간 이후부터 내동성이 증가했으며 5일째 가장 크게 증가하였다. 치사온도에서의 생존율도 저온처리한지 12시간 후부터 증가해서 5일 후 가장 높은 생존율을 보였다. 수용성당 함량은 저온순화 한지 1일 후부터 크게 증가했다. 몇몇 주요한 당 함량에서 볼 수 있듯이 포도당(Glucose)은 저온처리된 배추묘에서 함량이 가장 많은 당 성분으로 저온처리된 배추묘와 처리되지 않은 배추묘 사이에서 1일 후부터 계속 큰 차이를 나타냈으며 과당(Fructose)도 포도당과 비슷한 경향을 보였다. 저온처리를 시작할 때 자당(Sucrose)이 가장 많이 증가하는 경향을 보였고 처리 9시간 이후부터 큰 차이를 나타내기 시작했다. 당의 정량분석결과 배추묘에서 주요한 당 성분은 포도당이었으나, 저온처리시 크게 증가한 당 성분은 자당이었다. 최대내동성에 도달하기까지 5일이 걸렸으나 탈순화시 9시간후부터 생존율이 서서히 감소해 24시간만에 저온순화로 획득된 내동성이 모두 상실되었다. 전체 RNA와 수용성 단백질 함량 역시 24시간 경과 후 현저히 감소하였다.

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How to Explain the Contradiction of microRNA 200c Expression and Survival in Solid Tumors?: a Meta-analysis

  • Wang, Hui-Yu;Shen, Jie;Jiang, Chun-Ping;Liu, Bao-Rui
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권8호
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    • pp.3687-3690
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    • 2014
  • MicroRNA 200c is a microRNA 200 family member that plays an important role in regulation of the epithelial-to-mesenchymal transition (EMT). The prognostic value of microRNA 200c in solid tumors remains controversial because of inconsistent data. Here, we report a meta-analysis of the association of microRNA 200c expression and survival in patients with solid tumors. Pubmed was searched up to November 2013 for studies investigating microRNA 200c expression and overall survival (OS) in solid tumors. Hazard ratios (HRs) with 95% confidence intervals (CIs) for OS were extracted from each study. Pooled HR and CIs were calculated using the Mantel-Haenszel fixed-effects models. A total of five studies evaluating colorectal cancer, gastric cancer, ovarian cancer, pancreatic cancer and endometrial cancer were included in the analysis. Data were divided into tissue microRNA 200c expression group and serum microRNA 200c expression group. The combined HRs [95%CIs] estimated for OS were 0.62 [0.42-0.91] and 2.16 [1.32-3.52] respectively. Low expression of microRNA 200c in tumor tissue and high expression of microRNA 200c in serum are associated with worse survival in solid tumors. Further study is needed to elucidate this contradiction.

Ginsenoside Rh2 reduces m6A RNA methylation in cancer via the KIF26B-SRF positive feedback loop

  • Hu, Chunmei;Yang, Linhan;Wang, Yi;Zhou, Shijie;Luo, Jing;Gu, Yi
    • Journal of Ginseng Research
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    • 제45권6호
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    • pp.734-743
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    • 2021
  • Background: The underlying mechanisms of the potential tumor-suppressive effects of ginsenoside Rh2 are complex. N6-methyladenosine (m6A) RNA methylation is usually dysregulated in cancer. This study explored the regulatory effect of ginsenoside Rh2 on m6A RNA methylation in cancer. Methods: m6A RNA quantification and gene-specific m6A RIP-qPCR assays were applied to assess total and gene-specific m6A RNA levels. Co-immunoprecipitation, fractionation western blotting, and immunofluorescence staining were performed to detect protein interactions and distribution. QRT-PCR, dual-luciferase, and ChIP-qPCR assays were conducted to check the transcriptional regulation. Results: Ginsenoside Rh2 reduces m6A RNA methylation and KIF26B expression in a dose-dependent manner in some cancers. KIF26B interacts with ZC3H13 and CBLL1 in the cytoplasm of cancer cells and enhances their nuclear distribution. KIF26B inhibition reduces m6A RNA methylation level in cancer cells. SRF bound to the KIF26B promoter and activated its transcription. SRF mRNA m6A abundance significantly decreased upon KIF26B silencing. SRF knockdown suppressed cancer cell proliferation and growth both in vitro and in vivo, the effect of which was partly rescued by KIF26B overexpression. Conclusion: ginsenoside Rh2 reduces m6A RNA methylation via downregulating KIF26B expression in some cancer cells. KIF26B elevates m6A RNA methylation via enhancing ZC3H13/CBLL1 nuclear localization. KIF26B-SRF forms a positive feedback loop facilitating tumor growth.

Screening and functional validation of lipid metabolism-related lncRNA-46546 based on the transcriptome analysis of early embryonic muscle tissue in chicken

  • Ruonan, Chen;Kai, Liao;Herong, Liao;Li, Zhang;Haixuan, Zhao;Jie, Sun
    • Animal Bioscience
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    • 제36권2호
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    • pp.175-190
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    • 2023
  • Objective: The study was conducted to screen differentially expressed long noncoding RNA (lncRNA) in chickens by high-throughput sequencing and explore its mechanism of action on intramuscular fat deposition. Methods: Herein, Rose crown and Cbb broiler chicken embryo breast and leg muscle lncRNA and mRNA expression profiles were constructed by RNA sequencing. A total of 96 and 42 differentially expressed lncRNAs were obtained in Rose crown vs Cobb broiler chicken breast and leg muscle, respectively. lncRNA-ENSGALT00000046546, with high interspecific variability and a potential regulatory role in lipid metabolism, and its predicted downstream target gene 1-acylglycerol-3-phosphate-O-acyltransferase 2 (AGPAT2), were selected for further study on the preadipocytes. Results: lncRNA-46546 overexpression in chicken preadipocyte 2 cells significantly increased (p<0.01) the expression levels of AGPAT2 and its downstream genes diacylglycerol acyltransferase 1 and diacylglycerol acyltransferase 2 and those of the fat metabolism-related genes peroxisome proliferator-activated receptor γ, CCAAT/enhancer binding protein α, fatty acid synthase, sterol regulatory element-binding transcription factor 1, and fatty acid binding protein 4. The lipid droplet concentration was higher in the overexpression group than in the control cells, and the triglyceride content in cells and medium was also significantly increased (p<0.01). Conclusion: This study preliminarily concludes that lncRNA-46546 may promote intramuscular fat deposition in chickens, laying a foundation for the study of lncRNAs in chicken early embryonic development and fat deposition.

Molecular Cloning and mRNA Expression of the Porcine Insulin-responsive Glucose Transporter (GLUT4)

  • Zuo, Jianjun;Dai, Fawen;Feng, Dingyuan;Cao, Qingyun;Ye, Hui;Dong, Zemin;Xia, Weiguang
    • Asian-Australasian Journal of Animal Sciences
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    • 제23권5호
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    • pp.640-648
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    • 2010
  • Insulin-responsive glucose transporter 4 (GLUT4) is a member of the glucose transporter family and mainly presents in skeletal muscle and adipose tissue. To clarify the molecular structure of porcine GLUT4, RACE was used to clone its cDNA. Several cDNA clones corresponding to different regions of GLUT4 were obtained by amplifying reverse-transcriptase products of total RNA extracted from Landrace porcine skeletal muscles. Nucleotide sequence analysis of the cDNA clones revealed that porcine GLUT4 cDNA was composed of 2,491 base pairs with a coding region of 509 amino acids. The deduced amino acid sequence was over 90% identical to human, rabbit and cattle GLUT4. The tissue distribution of GLUT4 was also examined by Real-time RT-PCR. The mRNA expression abundance of GLUT4 was heart>liver, skeletal muscle and brain>lung, kidney and intestine. The developmental expression of GLUT4 and insulin receptor (IR) was also examined by Real-time RT-PCR using total RNA extracted from longissimus dorsi (LM), semimembranosus (SM), and semitendinosus (SD) muscle of Landrace at the age of 1, 7, 30, 60 and 90 d. It was shown that there was significant difference in the mRNA expression level of GLUT4 in skeletal muscles of Landrace at different ages (p<0.05). The mRNA expression level of IR also showed significant difference at different ages (p<0.05). The developmental change in the mRNA expression abundance of GLUT4 was similar to that in IR, and both showed a higher level at birth and 30 d than at other ages. However, there was no significant tissue difference in the mRNA expression of GLUT4 or IR (p>0.05). These results showed that the nucleotide sequence of the cDNA clones was highly identical with human, rabbit and cattle GLUT4 and the developmental change of GLUT4 mRNA in skeletal muscles was similar to that of IR, suggesting that porcine GLUT4 might be an insulin-responsive glucose transporter. Moreover, the tissue distribution of GLUT4 mRNA showed that GLUT4 might be an important nutritional transporter in porcine skeletal muscles.

생쥐 난소의 발생단계에 따른 Growth Differentiation Factor-9의 유전자 발현 (Expression of Growth Differentiation Factor-9 in the Mouse Ovaries at Different Developmental Stages)

  • 윤세진;이경아;고정재;차광열
    • 한국발생생물학회지:발생과생식
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    • 제3권1호
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    • pp.95-100
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    • 1999
  • Growth/differentiation factor-9 (GDF-9)은 transforming growth factor $\beta$ (TGF-$\beta$) superfamily의 member로서 난소의 난자에서만 특이적으로 발현되며 정상적인 난포발달에 있어 필수적인 성숙인자로 최근에 알려졌다. 본 연구는 RT-PCR을 통해 생쥐의 원시난포에서의 GDF-9 mRNA의 발현 여부와 함께 난포의 발달단계에 따른 상대적인 발현량을 분석하고자 실시하였다. 본 실험에는 ICR 생쥐를 사용하여 질전 (vaginal plug)이 확인된 날을 1일로 하여 임신 19일의 태아와 태어난 날을 1일로 하여 생후 1일, 10일, 21일, 28일된 생쥐 난소를 실험에 사용하였다. 각 발달단계의 난소조직으로부터 total RNA를 추출하여 GDF-9 유전자 발현 여부를 확인하였으며 이들을 $\beta$-actin에 대해 상대적인 정량분석을 하였다. GDF-9 유전자 발현은 아직은 성장을 시작하지 않은 임신 19일의 태아의 난소, 대부분이 원시난포로 이루어진 태어난 날의 생쥐 난소에서도 확인되었으며, 성장이 왕성하게 진행되고 있는 난포 즉, antrum 형성 이전의 growing follicles이 주를 이루는 생후 10일째의 난소에서 가장 높은 GDF-9 유전자 발현이 관찰되었다. 나머지 단계의 난소에서는 거의 비슷한 정도로 발현함을 관찰할 수 있었다. 본 연구의 결과는 생쥐의 원시난포에도 GDF-9 transcript가 존재한다는 것을 확실하게 증명하였으며, GDF-9이 생쥐의 초기 난포발달에 중요한 역할을 할 것이라는 가능성을 시사한다.

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Differential MicroRNA Expression Between Gastric Cancer Tissue and Non-cancerous Gastric Mucosa According to Helicobacter pylori Status

  • Lee, Jung Won;Kim, Nayoung;Park, Ji Hyun;Kim, Hee Jin;Chang, Hyun;Kim, Jung Min;Kim, Jin-Wook;Lee, Dong Ho
    • Journal of Cancer Prevention
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    • 제22권1호
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    • pp.33-39
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    • 2017
  • Background: MicroRNAs (miRNAs) are key post-translational mechanisms which can regulate gene expression in gastric carcinogenesis. To identify miRNAs responsible for gastric carcinogenesis, we compared expression levels of miRNAs between gastric cancer tissue and non-cancerous gastric mucosa according to Helicobacter pylori status. Methods: Total RNA was extracted from the cancerous regions of formalin-fixed, paraffin-embedded tissues of H. pylori-positive (n = 8) or H. pylori-negative (n = 8) patients with an intestinal type of gastric cancer. RNA expression was analyzed using a 3,523 miRNA profiling microarray based on the Sanger miRBase. Validation analysis was performed using TaqMan miRNA assays for biopsy samples from 107 patients consisted of control and gastric cancer with or without H. pylori. And then, expression levels of miRNAs were compared according to subgroups. Results: A total of 156 miRNAs in the aberrant miRNA profiles across the miRNA microarray showed differential expression (at least a 2-fold change, P < 0.05) in cancer tissue, compared to noncancerous mucosa in both of H. pylori-negative and -positive samples. After 10 promising miRNAs were selected, validations by TaqMan miRNA assays confirmed that two miRNAs (hsa-miR-135b-5p and hsa-miR-196a-5p) were significantly increased and one miRNA (hsa-miR-145-5p) decreased in cancer tissue compared to non-cancerous gastric mucosa at H. pylori-negative group. For H. pylori-positive group, three miRNAs (hsa-miR-18a-5p, hsa-miR-135b-5p, and hsa-miR-196a-5p) were increased in cancer tissue. hsa-miR-135b-5p and hsa-miR-196a-5p were increased in gastric cancer in both of H. pylori-negative and -positive. Conclusions: miRNA expression of the gastric cancer implies that different but partially common gastric cancer carcinogenic mechanisms might exist according to H. pylori status.

팔딱이 지렁이(Perionyx excavatus) DDX3 유전자의 동정 및 특성 (Identification and characteristics of DDX3 gene in the earthworm, Perionyx excavatus)

  • 박상길;배윤환;박순철
    • 유기물자원화
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    • 제23권1호
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    • pp.70-81
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    • 2015
  • Helicase는 NTP 결합의 화학적 에너지를 이용하여 이중가닥의 DNA와 RNA를 단일가닥으로 분해하여 다양한 생체반응에 기여하는 단백질로 알려져 있으며, 이 중 DEAD-box의 단백질은 주로 RNA와 관련된 대부분의 생화학적 반응에 작용하는 ATP 의존성 helicase로 알려져 있다. 또한 이 단백질 부류에 속하는 DEAD-box3 (DDX3) gene은 척추동물뿐만 아니라 무척추동물에서의 유성 생식과 무성 생식에서 생식세포 발달 및 재생과정 중 줄기세포 분화에 중요한 역할을 하는 인자로 알려져 있다. 이에 본 연구는 강한 재생능력을 가진 것으로 알려져 있는 팔딱이 지렁이(Perionyx excavatus)에서 DDX3 gene을 동정하고 그 발현양상을 알아보고자 환대를 포함하는 성체 지렁이의 두부를 절단하여 total RNA를 추출하고, 이를 주형으로 RT-PCR을 수행하여 full length의 DDX3 gene인 Pe-DDX3를 검출하였다. Pe-DDX3는 607개 아미노산 서열로 이루어져 있으며, DEAD-box 단백질 그룹 내에서 특이적으로 보존되어 있는 9개의 motif가 존재하고 있다. 다른 분류군에 속하는 동물들과의 multiple alignment를 통해 서열 내에 보존되어 있는 아미노산 서열을 확인할 수 있었으며, 아미노산 차원에서의 계통수 분석을 통해 DDX3 (PL10) 하부그룹에 속하는 것을 알 수 있었으며, 또한, 같은 그룹에 속하는 동물 중 P. dumerilii의 PL10a, b 단백질과 가장 가까운 유연관계를 확인 할 수 있었다.

약독 담배모자이크바이러스 II. RNA 및 외피단백질의 특성 (Studies on Mild Mutants of Tobacco Mosaic Virus II. Biochemical Properties of Ribonucleic Acid and Coat Protein)

  • 최장경;박원목
    • 한국식물병리학회지
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    • 제2권2호
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    • pp.121-128
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    • 1986
  • 액독 TMV, Tw 333 RNA 및 외피단백질에 대한 생화학적 특성을 조사하였다. Tw333-RNA는 $2.03\times10^6$dalton의 분자량을 나타냈고, 고기조성은 guanine 25.4, adenine 29.2, cytosine 17.5, uracil 27.9mol이었다. 열처리에서 얻어진 이 RNA의 농색효과는 $25.1\%$를 나타냈고, 이때 Tm치는 $47^{\circ}C$였다. 한편 Tw 333의 외피단백질은 17,500 dalton의 분자량을 보였으며, 16종의 아미노산으로 구성된 158개의 아미노산잔기를 나타냈다. Trypsin으로 분해한 단백질은 9종의 ninhydrin 양성반응 peptide를 형성하였다. 이들 약독 TMV, Tw333-RNA 및 외피단백질의 생화학적 특성은 원주 OM계통과 전반적으로 매우 유사하였다. 그러나 고기조성, 농색효과, 아미노산조성 및 peptide map에서 약간의 차가 인정되었다.

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Cytokine mRNA Expression in the Small Intestine of Weanling Pigs Fed Diets Supplemented with Specialized Protein or Peptide Sources

  • Zhao, J.;Harper, A.F.;Webb, K.E. Jr.;Kuehn, L.A.;Gilbert, E.;Xiao, X.;Wong, E.A.
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권12호
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    • pp.1800-1806
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    • 2008
  • Cytokines play a central role in the mucosal immune response and are involved in regulation of nutrient absorption, metabolism and animal growth. This study investigated the effect of diet manipulation with specialized protein or peptide sources on expression of cytokine (IL-1, IL-6, IL-10, and TNF-${\alpha}$) mRNA abundance in different intestinal regions and at different ages post-weaning in piglets. A total of 48 (17 days of age, $6.16{\pm}0.34kg\;BW$) weanling pigs were fed either a corn-soy/whey protein basal diet, the basal diet supplemented with spray-dried plasma protein (SDPP), or the basal diet supplemented with $Peptiva^{(R)}$, a hydrolyzed marine plant protein. A fourth treatment group was fed the SDPP diet, but the feed intake level was limited (SDPP-LF). Pigs were killed at 3 and 10 d, and intestinal cytokine mRNA was measured by real-time PCR using the relative quantification method. The SDPP-LF group exhibited an increased TNF-${\alpha}$ mRNA abundance compared with the ad libitum SDPP group (p<0.05). The TNF-${\alpha}$ and IL-10 mRNA abundance increased from the proximal to distal part of the intestine, and the mRNA abundance was greater (p<0.01) in the distal intestine as compared with the proximal and middle intestine. The cytokines IL-1-${\beta}$, IL-10 and TNF-${\alpha}$ mRNA abundance also increased from d3 to d10 postweaning (p<0.01). In summary, restricted feeding increased the TNF-${\alpha}$ mRNA abundance in the small intestine, however neither SDPP nor peptide supplementation affected cytokine mRNA expression. Abundance of mRNA for most cytokines examined in this study increased with age post-weaning, suggesting that during 10 d after weaning the mucosal immune system is still under development.