• Asian-Australasian Journal of Animal Sciences
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• 제31권6호
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• pp.812-819
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• 2018

Objective: An experiment was conducted to identify and characterize the circular RNA expression and metabolic characteristics in the liver of Jinhua pigs and Landrace pigs. Methods: Three Jinhua pigs and three Landrace pigs respectively at 70-day were slaughtered to collect the liver tissue samples. Immediately after slaughter, blood samples were taken to detect serum biochemical indicators. Total RNA extracted from liver tissue samples were used to prepare the library and then sequence on HiSeq 2500. Bioinformatic methods were employed to analyze sequence data to identify the circRNAs and predict the potential roles of differentially expressed circRNAs between the two breeds. Results: Significant differences in physiological and biochemical traits were observed between growing Jinhua and Landrace pigs. We identified 84,864 circRNA candidates in two breeds and 366 circRNAs were detected as significantly differentially expressed. Their host genes are involved in lipid biosynthetic and metabolic processes according to the gene ontology analysis and associated with metabolic pathways. Conclusion: Our research represents the first description of circRNA profiles in the porcine liver from two divergent phenotype pigs. The predicted miRNA-circRNA interaction provides important basis for miRNA-circRNA relationships in the porcine liver. These data expand the repertories of porcine circRNA and are conducive to understanding the possible molecular mechanisms involved in miRNA and circRNA. Our study provides basic data for further research of the biological functions of circRNAs in the porcine liver.

• 미생물학회지
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• 제47권4호
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• pp.289-294
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• 2011

Bacillus subtilis에 존재하는 DEAD-box RNA helicase 유전자의 결손이 저온 충격에 민감성을 보이는지를 조사하였다. 저온 충격에 민감한지를 알아 보기 위하여 대수기에($O.D_{600}$=0.5-0.6) 있는 세포를 $15^{\circ}C$도 낮추어 저온충격을 가하여 생장하는 정도를 조사하였다. DEAD-box RNA helicase 유전자 ydbR, yfmL, yqfR, deaD의 결손 균주들이 저온충격을 가하였을 때 ydbR 결손 균주의 생장이 야생형 균주에 비해 5배 정도 현저히 감소하였으나, 다른 DEAD-box RNA helicase 유전자의 (yfmL, yqfR, deaD) 결손은 야생형 균주와 비슷한 생장을 보였다. 저온에서의 유전자 발현을 알아보기 위하여 Northern blot으로 mRNA 양을 알아본 결과 $37^{\circ}C$에 비해 $15^{\circ}C$에서 ydbR과 yqfR의 mRNA전사물 증가를 확인할 수 있었고, 반면에 yfmL과 deaD의 전사 증가는 관찰되지 않았다. $37^{\circ}C$에서 $15^{\circ}C$로 저온 충격을 가하면 ydbR mRNA 양의 뚜렷한 증가를 확인하였고, 전사 억제제인 rifampicin를 처리하여 ydbR mRNA의 양을 조사하였을 때 $15^{\circ}C$ 조건에서는 mRNA 양이 거의 유지하는 반면에 $37^{\circ}C$ 조건에서는 급격한 mRNA의 감소가 일어나 전사과정에서 유도되기 보다는 전사 후 전사물의 안정에 기인하는 것으로 보인다. 이와 관련하여 ydbR 유전자의 5' UTR (untranaslated region) 부근에서 csp (cold shock protein) 유전자에서 관찰되는 cold box element를 확인하였고, ydbR이 저온 충격 조건에서 발현되는 과정이 csp와 유사하게 전사물의 안정성에 기인함을 알 수 있었다.

• Animal Bioscience
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• 제36권4호
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• pp.555-569
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• 2023

Objective: The objective of this study was to investigate the effects of N⁶-Methyladenosine modification-circRNA-zinc finger protein 638 (m⁶A-circRNA-ZNF638) on the induced activation of secondary hair follicle (SHF) stem cells with its potential mechanisms in cashmere goats. Methods: The m⁶A modification of ZNF638 was analyzed using methylation immunoprecipitation with real-time quantitative polymerase chain reaction technique in SHF stem cells. The effects of circRNA-ZNF638 on the induced activation of SHF stem cells in m⁶A dependence were evaluated through the overexpression of circRNA-ZNF638/its m⁶A-deficient mutants in circRNA-ZNF638 knockdown SHF stem cells. The competitive binding of miR-361-5p to circRNA-ZNF638/Wnt5a 3'- untranslated region was analyzed through Dual-luciferase reporter assay. Results: The m⁶A-circRNA-ZNF638 had significantly higher transcription at anagen SHF bulge of cashmere goats compared with that at telogen, as well as it positively regulated the induced activation of SHF-stem cells in cashmere goats. Mechanismly, m⁶A-circRNA-ZNF638 sponged miR-361-5p to heighten the transcriptional expression of Wnt5a gene in SHF-stem cells. We further demonstrated that the internal m⁶A modification within circRNA-ZNF638 is required for mediating the miR-361-5p/Wnt5a pathway to regulate the induced activation of SHF stem cells through an introducing of m⁶A-deficient mutant of circRNA-ZNF638. Conclusion: The circRNA-ZNF638 contributes the proper induced activation of SHF-stem cells in cashmere goats in m⁶A-dependent manner through miR-361-5p/Wnt5a axis.

• 한국식물병리학회지
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• 제3권4호
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• pp.239-244
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• 1987

감자 걀쪽바이로이드(PSTV) RNA분자를 포함한 저분자량의 식물성 RNA분자에 대해서 0M부터 8M까지의 요소농도구배를 가진 폴리아크릴아마이드겔을 이용하여 1/40, 1/10로 희석한 완충액조건과 $17^{\circ}C,\;37^{\circ}C,\;57^{\circ}C$의 온도조건에서 전기영동을 실시하였다. 바이로이드의 RNA분자가 환상과 선상의 두가지 분자로 나뉘어지는 변성조건에서는 1/40로 희석한 TBE완충액을 포함하는 요소농도구배겔을 $57^{\circ}$에서 전기영동을 실시하는 것이 효과적이었다. 변성된 환상의 바이로이드분자에 대한 전기 영동적 이동성은 주로 요소의 농도에 따라 영향을 받는 것으로 보이는데, 이와 더불어 저 농도의 TBE완충액이 요소농도구배겔에서 환상과 선상바이로이드분자 사이에서 전기영동적 분리를 증가시켜 주었다.

• 환경생물
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• 제33권4호
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• pp.433-440
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• 2015

The Apostichopus japonicus is an important species in some Asia countries including Korea, China and Japan. The purpose of the present study was to investigate the differential gene expression of heat shock protein90 (Hsp90) and ferritin as a biomarker for the thermal stress during water temperature rising in the sea cucumber, A. japonicus. The A. japonicus (1.4 g) was cultured in incubator of separate temperature ($15^{\circ}C$, $20^{\circ}C$, $25^{\circ}C$ and $30^{\circ}C$) for each 0, 3, 6, 12, 24, 48 hours. The mRNA expression levels of Hsp90 and ferritin were examined using RT-PCR assay. Results showed that, the expression of Hsp90 mRNA was not significantly changed at $15^{\circ}C$. The expression of Hsp90 mRNA was significantly increased at high temperature such as $20^{\circ}C$ and $25^{\circ}C$. Furthermore, Hsp90 mRNA was early increased at $25^{\circ}C$ than $20^{\circ}C$. The ferritin mRNA was similar expression pattern with Hsp90. But, Hsp90 mRNA was more sensitive than ferritin mRNA at high thermal stress. These results indicate that Hsp90 and ferritin mRNAs were involved in the temperature changes response and may be play an important role in mediating the thermal stress in A. japonicas.

• Applied Biological Chemistry
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• 제38권6호
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• pp.522-527
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• 1995

흑조위축병 바이러스(RBSDV)에 대한 antiviral agent를 개발하기위하여 RBSDV 게놈 조각 3번을 절단하는 망치머리형 구조의 라이보자임을 설계하였다. 라이보자임과 기질의 oligonucleotides는 DNA 합성기로 합성 후 서로 합치고, pBluescript KS(+)에 삽입하였다. 라이보자임과 기질 RNA는 $T_3$ RNA polymerase로 기내 전사하여 각각 193과 183 nucleotide의 RNA를 얻었다. 기질 RNA는 라이보자임 RNA에 의해 $55^{\circ}C$에서 2개의 조각으로 절단되었으나 $37^{\circ}C$에서는 절단되지 않았다. 또한 RBSDV의 3번 조각 RNA도 동일한 라이보자임에 의해 절단되었다. 이러한 결과로 합성된 헤머해드 라이보자임은 기내 절단 능력을 가지며 형질전환 식물체에서 antiviral agent로 이용될 수 있을 것이다.

• Journal of Gastric Cancer
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• 제20권3호
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• pp.300-312
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• 2020

Purpose: Circular RNAs (circRNAs) are a new class of RNA molecules whose function is largely unknown. There is a growing evidence that circRNAs play an important regulatory role in the progression of a variety of human cancers. However, the exact roles and the mechanisms of circRNAs in gastric cancer are not clear. In this study, we aimed to elucidate the mechanism of hsa_circ_0005556. Materials and Methods: Real-time quantitative polymerase chain reaction was used to detect the expression of hsa_circ_0005556, miR-4270, and matrix metalloproteinase-19 (MMP19) in gastric cancer tissues and cell lines. The expression of hsa_circ_0005556 in gastric cancer cells was silenced by lentivirus, and cell proliferation, invasion, migration, and tumorigenesis in nude mice were assessed to evaluate the function of hsa_circ_0005556 in gastric cancer. Results: The expression of hsa_circ_0005556 in gastric cancer tissues and gastric cancer cell lines was higher compared to normal controls. In vitro, the downregulation of hsa_ circ_0005556 significantly inhibited proliferation, migration, and invasion of gastric cancer cells. In vivo, the downregulation of hsa_circ_0005556 suppressed tumor growth in nude mice. Conclusions: Our study shows that the hsa_circ_0005556/miR-4270/MMP19 axis is involved in proliferation, migration, and invasion of gastric cancer cells through the competing endogenous RNA (ceRNA) mechanism.

• Applied Biological Chemistry
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• 제37권1호
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• pp.56-63
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• 1994

오이모자이크 바이러스(CMV)의 외피단백질 유전자의 일정한 염기서열을 보유하는 부분과 CMV RNA에 대항한 헤머헤드(hammerhead) 구조의 ribozyme을 만드는 올리고뉴클 레오타이드(oligonucleotide, nt)를 DNA 합성기를 이용하여 제조하였다. 올리고뉴클레오타이드의 양쪽가닥을 서로 합친 후 제한효소 BamHl과 SacI으로 처리하여 플라스미드 pBS SK(+)에 삽입하였고 CMV 기질과 ribozyme 클론의 염기서열을 결정하여 확인하였다. 기질과 ribozyme 클론 $1\;{\mu}g$ BssHII이나 SspI으로 처리한후 T7 RNA 합성효소를 이용하여 튜브내에서 전사반응을 실시하였다. 제한효소 BssHII를 처리한 경우 만들어진 기질 RNA의 크기는 176 nt 였는데 50 nt의 CMV RNA 염기, 6 nt의 Xbal 제한효소 염기, 120 nt의 벡터에서 비롯된 염기를 포함한다. Ribozyme RNA의 크기는 164 nt인데 38 nt의 ribozyme 염기부분과 그외는 기질의 것과 같은 염기를 포함한다. CMV 기질 RNA는 ribozyme RNA에 의하여 특이적으로 절단되어 96 nt와 80 nt 두개의 조각을 만들었다. 이러한 특이적 절반은$37^{\circ}C$ 보다 $55^{\circ}C$에서 더 빠르게 일어났다. SspI으로 처리한 경우 만들어진 기질 RNA(2234 nt)도 역시 위치 ribozyme에 의해 두조각으로 절반되었으며 SspI 처리 후 만들어진 ribozyme RNA(2222 nt)에 의해서 특이적으로 절단되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제31권10호
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• pp.1550-1557
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• 2018

Objective: Circular RNAs (circRNAs) are a newfound class of non-coding RNA in animals and plants. Recent studies have revealed that circRNAs play important roles in cell proliferation, differentiation, autophagy and apoptosis during development. However, there are few reports about muscle development-related circRNAs in livestock. Methods: RNA sequencing analysis was employed to identify and annotate circRNAs from longissimus dorsi of sheep. Reverse transcription followed by real-time quantitative (q) polymerase chain reaction (PCR) analysis verified the presence of these circRNAs. Targetscan7.0 and miRanda were used to analyse the interaction of circRNA-microRNA (miRNA). To investigate the function of circRNAs, an experiment was conducted to perform enrichment analysis hosting genes of circRNAs using gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathways. Results: About 75.5 million sequences were obtained from RNA libraries of sheep skeletal muscle. These sequences were mapped to 729 genes in the sheep reference genome. We identified 886 circRNAs, including numerous circular intronic RNAs and exonic circRNAs. Reverse transcription PCR (RT-PCR) and DNA sequencing analysis confirmed the presence of several circRNAs. Real-Time RT-PCR analysis exhibited resistance of sheep circRNAs to RNase R digestion. We found that many circRNAs interacted with muscle-specific miRNAs involved in growth and development of muscle, especially circ776. The GO and KEGG enrichment analysis showed that hosting genes of circRNAs was involved in muscle cell development and signaling pathway. Conclusion: The study provides comprehensive expression profiles of circRNAs in sheep skeletal muscle. Our study offers a large number of circRNAs to facilitate a better understanding of their roles in muscle growth. Meanwhile, we suggested that circ776 could be analyzed in future study.

• 한국식품과학회지
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• 제27권1호
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• pp.129-133
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• 1995

배양환경이 효모 RNA 축적과 autolysis 효율에 미치는 영향을 알아보기 위하여 연속배양을 실시하였다. Dilution rate의 증가에 따라 균체내의 RNA 함량은 증가하여 dilution rate=0.35 $h^{-1}$에서 14.8%를 나타내었고, 더불어 RNA productivity와 specific RNA productivity도 증가하였다. 고정된 D=0.2 $h^{-1}$에서 최적 균체량은 배양온도 $30^{\circ}C$ 에서 얻어졌으나, RNA 함량은 배양온도가 낮아질수록 높은 값을 보여 $24^{\circ}C$에서 12.7%로 나타났다. 한편, autolysis 효율 측면에서 균체 생육속도가 증가할수록 효모 extract의 추출율은 감소하였고, 효모 extract의 분해율은 증가하는 경향을 보였으나, 배양온도는 효모 autolysis 효율에 큰 영향을 미치지 않았다.