• BMB Reports
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• 제45권5호
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• pp.293-298
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• 2012

The p38 mitogen-activated protein kinase (MAPK) is involved in various processes, including stress responses, development, and differentiation. However, little information on p38 MAPK in insects is available. In this study, a p38 MAPK gene, $Accp38b$, was isolated from $Apis$ $cerana$ $cerana$ and characterized. The quantitative real-time PCR (Q-PCR) analysis revealed that $Accp38b$ was induced by multiple stressors. Notably, the expression of $Accp38b$ was relatively higher in the pupae phase than in other developmental phases. During the pupae phase, Accp38b expression was higher in the thorax than in the head and abdomen and higher in the fat body than in the muscle and midgut. Immunohistochemisty showed significant positive staining of Accp38b in sections from the brain, eyes, fat body, and midgut of $A.$ $cerana$ $cerana$. These results suggest that Accp38b may play a crucial role in stress responses and have multiple aspects function during development.

• Asian Pacific Journal of Cancer Prevention
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• 제15권15호
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• pp.6269-6273
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• 2014

Growing evidence suggests that miR-150-5p has an important role in regulating genesis of various types of cancer. However, the roles and the underlying mechanisms of miR-150-5p in development of colorectal cancer (CRC) remain largely unknown. Transwell chambers were used to analyze effects on cell migration and invasion by miR-150-5p. Quantitative real-time PCR (qRT-PCR), Western blotting and dual-luciferase 3' UTR reporter assay were carried out to identify the target genes of miR-150-5p. In our research, miR-150-5p suppressed CRC cell migration and invasion, and MUC4 was identified as a direct target gene. Its effects were partly blocked by re-expression of MUC4. In conclusiomn, miR-150-5p may suppress CRC metastasis through directly targeting MUC4, highlighting its potential as a novel agent for the treatment of CRC metastasis.

• ALGAE
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• 제32권3호
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• pp.235-244
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• 2017

Freshwater algae living in shallow waters have evolved various photomovement to stay in the optimum light condition for survival. Previous action-spectra investigations showed that Spirogyra filaments have phototropic movement in blue light. To decipher the genetic control of phototropic movement, two phototropin homologues were isolated from Spirogyra varians, and named SvphotA and SvphotB. Both phototropins have similar molecular structure consisted of two light-oxygen-voltage domains (LOV1, LOV2) and a serine / threonine kinase domain. SvphotA and SvphotB had 48.7% sequence identity. Phylogenetic analysis showed SvphotA and SvphotB belong to different clades suggesting early divergence, possibly before the divergence of land plants from the Zygnematales. Quantitative PCR and northern blot analysis showed that SvphotA and SvphotB responded differently to red and blue light. SvphotA was consistently expressed in the dark and in blue light, while SvphotB was expressed only when the plants were exposed to light. When the filaments were exposed to red light, SvphotA was significantly downregulated whereas SvphotB was highly upregulated. These results suggest that the two phototropins may have different roles in the photoresponse in S. varians.

• 대한의생명과학회지
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• 제26권1호
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• pp.8-13
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• 2020

Cervical cancer is the fourth most common cancer in women, with approximately 528,000 new cases and 266,000 women dying of it per year in the world. MicroRNAs have recently been in the spotlight as potential biomarkers that regulate gene expression and are involved in tumorigenesis. In the present study, we evaluated miR-1260b as a potential biomarker for screening of cervical cancer by quantitative reverse transcription PCR. We profiled the TCGA data of miR-1260b in 307 cervical cancer tissues. Then, miR-1260b expression levels in 10 cervical cancer tissues and 10 noncancerous tissues were investigated in a pilot study. miR-1260b was found to be significantly up-regulated in cervical cancer FFPE tissues as compared to those in cervical normal FFPE tissues (P = 0.006). The mean expression level of miR-1260b in late-stage (IIB-IVB) was higher than in those with early-stage (IA-IIA). Furthermore, high miR-1260b was found to be associated with high hTERT and Ki-67 mRNA expression, which are representative of tumor prognosis. The results of the pilot study suggest that miR-1260b may be used as a novel biomarker for improving the diagnosis of cervical cancer.

• 한국작물학회지
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• 제66권4호
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• pp.365-374
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• 2021

이소플라본 함량이 높으며 장류콩인 대풍2호와 이소플라본 함량이 낮으며 나물콩인 풍산나물콩을 재료로 하여 콩 발아기간 동안 이소플라본 생합성 관련 유전자 6종(CHS6, HID, IF7GT, IF7MaT, GmIMaT1 및 GmIMaT3)의 발현량을 qRT-PCR로 분석하였다. 1. 공시재료 모두 발아기간 경과에 따라 이소플라본 함량이 높아졌으며, 총 이소플라본 함량 중 malonyl-glucosides 함량이 80% 이상을 차지하여 제일 높았으며 acetyl-glucosides는 거의 분석되지 않았다. 한편, 자엽과 배축에서 이소플라본 축적 정도가 각각 다르게 나타났으며 개별 이소플라본 함량에서도 차이가 있었다. 2. 이소플라본 생합성 관련 유전자들은 콩 발아시기 경과에 따라 발현량이 높아져 이소플라본 축적 정도와 상관이 있는 것으로 판단되나 유전자들의 발현량이 시기별로 각각 달라 개별 이소플라본 함량과 유전자간 뚜렷한 상관은 찾을 수 없었다. 3. HID 유전자는 대풍2호와 풍산나물콩 모두 발아 3일차를 제외하고는 발아시간 경과에 따라 유전자의 발현량이 높아졌으나 다른 유전자들의 상대적 발현량은 품종 간 차이가 있었으며, 또한 발아시간 경과에 따른 유전자의 발현 양상은 유전자별로 각각 달랐다. 4. 발아시간 별로 유전자들의 상대적 발현량을 비교한 결과 품종 간 차이가 있었으며 자엽과 배축에서도 상대적 발현량이 다르게 나타났다. 자엽에서는 GmIMaT1을 제외한 다른 유전자들의 발현량이 대풍2호와 풍산나물콩에서 비슷하게 나타난 반면, 배축에서는 HID 발현량이 2품종 모두 높게 나타났으나 다른 유전자들은 유전자 발현량에 일정한 경향이 없었다.

• 생명과학회지
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• 제30권12호
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• pp.1070-1077
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• 2020

암세포에 항암제를 처리하게 되면, 세포증식, 이동성 또는 약물 내성과 관련된 많은 유전자들의 발현 변화가 발생하며, 유전자 발현 변화는 상호간의 조절 네트워크에 의해 밀접하게 연결될 수도 있다고 추측된다. 본 연구에서 p53 유전자 유무가 다른 A549와 H1299 인간 폐암세포에 독소루비신을 처리하면, 원종양유전자인 FosB의 발현은 증가하지만, 원종양유전자인 SETDB1의 발현은 감소하지만, 단백질 발현의 양적인 차이가 발생한다는 사실을 알 수 있었다. TF motif binding 분석 프로그램을 이용하여 SETDB1과 FosB 프로모터지역에서의 p53단백질의 결합가능성을 분석한 결과, SETDB1의 경우 18부위, FosB의 경우 21 부위의 p53 결합부위를 예측할 수 있었다. SETDB1과 FosB 프로모터의 subcloning하여 luciferase 분석을 수행한 결과, p53은 SETDB1과 FosB을 음성적으로 조절한다는 사실을 알 수 있었다. 또한, H1299 세포에 p53의 과발현은 SETDB1 과 FosB의 발현을 감소시킬 수 있음을 RT-PCR, western blot, qPCR, 면역염색 실험을 통해 확인하였다. 이러한 결과를 종합하여 본다면, p53에 의한 SETDB1과 FosB 유전자 발현 조절은 항암제 처리과정에서 나타나는 암세포의 사멸과 생존에 대한 기능적 조절 네트워크로 사료된다.

• Mycobiology
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• 제46권1호
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• pp.64-71
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• 2018

The aim of this study was to investigate the pattern of distribution of mating type (MAT) genes of Tuber indicum in ectomycorhizosphere soils from natural T. indicum-producing areas and cultivated truffle orchards and ascocarp samples from different regions. Quantitative real-time PCR and multiplex PCR were used to weight the copy numbers of MAT1-1-1 and MAT1-2-1 in natural truffle soils and cultivated orchard soils. The effect of limestone on the pattern of truffle MAT genes and the correlation between soil properties and the proportion of MAT genes were also assessed. These results indicated that an uneven and nonrandom distribution of MAT genes was common in truffle-producing areas, cultivated truffle orchards, and ascocarps gleba. The competition between the two mating type genes and the expansion of unbalanced distribution was found to be closely related to truffle fructification. Limestone treatments failed to alter the proportion of the two mating type genes in the soil. The content of available phosphorus in soil was significantly correlated with the value of MAT1-1-1/MAT1-2-1 in cultivated and natural ectomycorhizosphere soils. The application of real-time quantitative PCR can provide reference for monitoring the dynamic changes of mating type genes in soil. This study investigates the distributional pattern of T. indicum MAT genes in the ectomycorhizosphere soil and ascocarp gleba from different regions, which may provide a foundation for the cultivation of T. indicum.

• Asian-Australasian Journal of Animal Sciences
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• 제32권2호
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• pp.170-175
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• 2019

Objective: Uncoupling protein 3 gene (UCP3) is a candidate gene associated with the meat quality of pigs. The aim of this study was to explore the regulation mechanism of UCP3 expression and provide a theoretical basis for the research of the function of porcine UCP3 gene in meat quality. Methods: Bisulfite sequencing polymerase chain reaction (PCR) and quantitative real-time PCR (Q-PCR) were used to analyze the methylation of UCP3 5′-flanking region and UCP3 mRNA expression in the adipose tissue or skeletal muscle of three pig breeds at different ages (1, 90, 210-day-old Putian Black pig; 90-day-old Duroc; and 90-day-old Dupu). Results: Results showed that two cytosine-guanine dinucleotide (CpG) islands are present in the promoter region of porcine UCP3 gene. The second CpG island located in the core promoter region contained 9 CpG sites. The methylation level of CpG island 2 was lower in the adipose tissue and skeletal muscle of 90-day-old Putian Black pigs compared with 1-day-old and 210-day-old Putian Black pigs, and the difference also existed in the skeletal muscle among the three 90-day-old pig breeds. Furthermore, the obvious changing difference of UCP3 mRNA expression was observed in the skeletal muscle of different groups. However, the difference of methylation status and expression level of UCP3 gene was not significant in the adipose tissue. Conclusion: Our data indicate that UCP3 mRNA expression level was associated with the methylation status of UCP3 promoter in the skeletal muscle of pigs.

• Mycobiology
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• 제47권1호
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• pp.59-65
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• 2019

Agrobacterium tumefaciens-mediated transformation (ATMT), as a simple and versatile method, achieves successful transformation in the yeast-like cells (YLCs) of Tremella fuciformis with lower efficiency. Establishment of a more efficient transformation system of YLCs is important for functional genomics research and biotechnological application. In this study, an enzymolysis-assisted ATMT method was developed. The degradation degree of YLCs depends on the concentration and digestion time of Lywallzyme. Lower concentration (${\leq}0.1%$) of Lywallzyme was capable of formation of limited wounds on the surface of YLCs and has less influence on their growth. In addition, there is no significant difference of YLCs growth among groups treated with 0.1% Lywallzyme for different time. The binary vector pGEH under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter was utilized to transform the enzymolytic wounded YLCs with different concentrations and digestion time. The results of PCR, Southern blot, quantitative real-time PCR (qRT-PCR) and fluorescence microscopy revealed that the T-DNA was integrated into the YLCs genome, suggesting an efficient enzymolysis-assisted ATMT method of YLCs was established. The highest transformation frequency reached 1200 transformants per $10^6$ YLCs by 0.05% (w/v) Lywallzyme digestion for 15 min, and the transformants were genetically stable. Compared with the mechanical wounding methods, enzymolytic wounding is thought to be a tender, safer and more effective method.

• The Plant Pathology Journal
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• 제38권6호
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• pp.656-664
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• 2022

Pectobacterium odoriferum is the primary causative agent in Kimchi cabbage soft-rot diseases. The pathogenic bacteria Pectobacterium genera are responsible for significant yield losses in crops. However, P. odoriferum shares a vast range of hosts with P. carotovorum, P. versatile, and P. brasiliense, and has similar biochemical, phenotypic, and genetic characteristics to these species. Therefore, it is essential to develop a P. odoriferumspecific diagnostic method for soft-rot disease because of the complicated diagnostic process and management as described above. Therefore, in this study, to select P. odoriferum-specific genes, species-specific genes were selected using the data of the P. odoriferum JK2.1 whole genome and similar bacterial species registered with NCBI. Thereafter, the specificity of the selected gene was tested through blast analysis. We identified novel species-specific genes to detect and quantify targeted P. odoriferum and designed specific primer sets targeting HAD family hydrolases. It was confirmed that the selected primer set formed a specific amplicon of 360 bp only in the DNA of P. odoriferum using 29 Pectobacterium species and related species. Furthermore, the population density of P. odoriferum can be estimated without genomic DNA extraction through SYBR Green-based real-time quantitative PCR using a primer set in plants. As a result, the newly developed diagnostic method enables rapid and accurate diagnosis and continuous monitoring of soft-rot disease in Kimchi cabbage without additional procedures from the plant tissue.