• The Plant Pathology Journal
• /
• 제27권1호
• /
• pp.33-36
• /
• 2011

The genome of a potyvirus isolate associated with chlorotic spots and vein banding symptoms on Spathiphyllum spp., in Andhra Pradesh state, India was amplified by RT-PCR using degenerate potyvirus primers, amplicons cloned, and sequence (1.6 kb) analyzed. This virus isolate shared maximum identity of 74.8% and 80.2% at coat protein (CP) gene nucleotide (906 nucleotides) and amino acid (302 amino acids) levels, respectively with Dasheen mosaic virus (DsMV)-M13 isolate reported from China. But its 3'-UTR (258 nucleotides) had maximum identity of 62.5% with DsMV-Vietnam isolate. The deduced molecular weight of CP is 33.57 kDa and it contained DAG triplet in its N-terminal region. In CP amino acid based phylogenetic analysis, this virus isolate represented a separate branch but closer to DsMV isolates cluster. Based on the molecular criteria set for the discrimination of species and genus in the Potyviridae family, the present virus isolate was identified as a distinct virus species in the genus Potyvirus and proposed the name Spathiphyllum chlorotic vein banding virus (SCVbV).

• Biomolecules & Therapeutics
• /
• 제27권1호
• /
• pp.78-84
• /
• 2019

Cell therapeutic agents for treating degenerative brain diseases using neural stem cells are actively being developed. However, few systems have been developed to monitor in real time whether the transplanted neural stem cells are actually differentiated into neurons. Therefore, it is necessary to develop a technology capable of specifically monitoring neuronal differentiation in vivo. In this study, we established a system that expresses cell membrane-targeting red fluorescent protein under control of the Synapsin promoter in order to specifically monitor differentiation from neural stem cells into neurons. In order to overcome the weak expression level of the tissue-specific promoter system, the partial 5' UTR sequence of Creb was added for efficient expression of the cell surface-specific antigen. This system was able to track functional neuronal differentiation of neural stem cells transplanted in vivo, which will help improve stem cell therapies.

• 원예과학기술지
• /
• 제28권4호
• /
• pp.648-655
• /
• 2010

본 연구에서는 들잔디와 돌연변이체에서 Glyphosate 내성 관련 유전자인 EPSPS를 코딩하는 cDNA를 분리하여, 시퀸스 비교분석과 발현 양상 차이 등에 관하여 조사하였다. 5'/3' RACE를 통하여 밝혀진 EPSPS의 cDNA는 각각 1176bp의 open reading frame으로 이루어져 있으며 391개의 아미노산을 코딩하고 있었다. 이는 보고된 다른 EPSPS 유전자들과 높은 유사성을 가지고 있다. Genomic southern 결과 들잔디 내에 EPSPS 유전자는 단일copy로 존재하였다. Wild type과 제초제 내성 변이체의 EPSPS 유전자는 6개의 아미노산 시퀀스의 차이를 보였으며 기존에 보고된 EPSPS active site에서 시퀀스의 차이를 나타냈다. 한편 glyphosate의 독성기작의 하나인 EPSPS 효소활성 저해 작용을 알아본 결과, 내성 개체에서 높은(1.5배 이상) EPSPS 활성을 확인할 수 있었다. Northern 분석과 RT-PCR로 제초제 처리 시간에 따른 EPSPS의 발현량을 살펴본 결과, 제초제 처리 후 시간이 경과할수록 발현량이 증가하다가 7일 이후에는 감소하였고, wild type에 비해 glyphosate 내성 선발개체에서 전체적인 발현량이 높음을 알 수 있었다. 본 연구 결과 선발된 glyphosate 내성 돌연변이체는 높은 EPSPS 효소활성 증가 및 EPSPS active site의 아미노산 시퀀스 변화를 통해 원할하고 지속적인 방향족 아미노산의 합성이 가능한 것으로 보여졌다. 본 실험을 통해 얻어진 glyphosate 내성 잔디 돌연변이체와 방법은 앞으로 유용한 제초제 저항성 돌연변이 품종개발 연구에 활용될 수 있을 것으로 기대된다.

• 식물조직배양학회지
• /
• 제28권5호
• /
• pp.231-237
• /
• 2001

Differential screening을 통해 Moriguchi등 (1998)이 분리한 유전자와 상동성을 나타내는 CitMT45 유전자의 cDNA를 분리하였다. 본 실험에서 분리한 cDNA는 Moriguchi등 (1998)이 분리한 cDNA에 비해 긴 3' UTR을 가지고 있었다. 잎과 과피, 과육에서 CitMT45 유전자의 발현분석을 northern blot을 통해 수행한 결과, 발달단계에 따라 증가하는 비슷한 앙상을 관찰할 수 있었으나, 과육, 과피, 잎의 순으로 그 발현 양이 많았다. 이들의 발현조절에 대한 정보를 얻기 위해 게놈 DNA를 분리한 결과, CitMT45 게놈 구조는 3개의 exon과 2개의 intron으로 구성되어 있었고, primer extension 분석을 통해 CitMT45 유전자의 발현은 3개의 부위에서 개시되고 있음을 알 수 있었다. 전사개시부위의 5'upstream 지역에서 TATA box와 CCAAT box뿐만 아니라, 금속이온과 온도변화에 의한 조절에 중요한 부위로 알려진 cis-element를 발견하였다.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권4호
• /
• pp.1553-1556
• /
• 2012

Introduction: Antimetabolites may cause severe toxicity and even toxic death in cancer patients. Our aim was to evaluate the relationship between antimetabolite toxicity and pharmacogenetics in patients with severe clinical toxicity or alanine transaminase (ALT) elevation after fluorouracil (5FU), capecitabine or methotrexate administration. Patients and Methods: Cancer patients with severe antimetabolite toxicity were evaluated for methylenetetrahydrofolate reductase (MTHFR) gene C667T, thymidilate synthase (TS) gene 5´UTR variable number of tandem repeats (VNTR), dihydroprymidine dehydrogenase (DPYD) gene IVS14+1G/A, Xeroderma pigmentosum (XPD) gene Lys751Gln and X-ray repair cross-complementing group 1 (XRCC1) gene Arg399Gln polymorphisms. Results: Eighteen patients were enrolled, with a male/female ratio of 0.8. They had osteosarcoma in methotrexate group (n=7), gastrointestinal malignancies in 5FU group (n=9) and breast cancer in the capecitabine group (n=2). Mucositis and dermatitis occurred in all groups, together with ALT elevation in the methotrexate group and 2 toxic deaths were encountered. DPYD, TS, MTHFR, XPD and XRCC1 gene polymorphism rare allele frequencies were observed to be higher than in the general population. Conclusion: Pharmacogenetics might contribute to tailored therapy.

• Asian Pacific Journal of Cancer Prevention
• /
• 제17권sup3호
• /
• pp.71-75
• /
• 2016

Pancreatic cancer is a leading cause of fatality worldwide. Several population studies have been conducted on genetic diagnosis of pancreatic cancer but the results from epidemiologic studies are very limited. CYP17A gene has a role in disease formation but its influence on pancreatic cancer is unclear. A polymorphism in the 5'UTR promoter region of CYP17A1-34T/C (A1/A2) has been associated with multiple cancers. The aim of the current study was to assess associations of this polymorphism and socio-demographic risk factors with pancreatic cancer. A total of 255 and 320 controls were enrolled in the study, and were genetically analyzed through PCR-RFLP. Statistical analysis was conducted with observed genotype frequencies and odds ratios (ORs) and 95% CIs were estimated using unconditional logistic regression. The impact of socio-demographic factors was accessed through Kaplen-Meir analysis. According to our results, the A2/A2 genotype was significantly associated with pancreatic cancer (OR=2.1, 95%CI = 1.3-3.5). Gender female (OR=2.6, 95%CI=1.8-3.7), age group 80s/80+ years (OR=2.2, 95% CI=1.2-4), smoking both former (OR=4.6, 95% CIs=2.5-8.8) and current (OR=3.6, 95% CI=2-6.7), and family history (OR=7.1; 95%CI = 4.6-11.4) were also found associated with increased risk. Current study suggests that along with established risk factors for pancreatic cancer CYP17A1-34T/C may play a role. However, on the basis of small sample size the argument cannot be fully endorsed and larger scale studies are recommended.

• Asian-Australasian Journal of Animal Sciences
• /
• 제19권2호
• /
• pp.165-170
• /
• 2006

Using the somatic cell hybrid panel (SCHP) and radiation hybrid (IMpRH) panel, porcine SOCS2 gene was mapped at SSC5 (1/2) q21-q24 and closely linked with SW1383 marker (47 cR in distance), while SOCS3 gene was assigned to SSC12p11-(2/3p13) and closely linked with SW2490 (43 cR). The reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to detect the expression of these two genes in the different tissues and the results showed that both SOCS2 and SOCS3 genes were widely expressed in tissues investigated (heart, liver, spleen, lung, kidney skeletal muscle, fat and brain), although some tissues showed lower gene expression. Moreover, SOCS2 and SOCS3 genes had different expression levels at different stages, in different tissues and in different breeds. A G/A substitution, which can be recognized by restriction enzyme of Cfr421, was observed in 5' untranslated region (5'-UTR) of SOCS2 gene. The allele frequencies was investigated by PCR-restriction fragment length polymorphism (PCR-RFLP) method and it showed that the allele frequency among Dahuabai, Erhualian, Yushan, Qingping, Large white and Landrace tested were different. Association analysis in a cross experimental populations revealed no significant association between the SOCS2 gene polymorphism and the economic traits investigated. The full-length coding regions (CDs) of porcine SOCS3 gene was obtained by RT-PCR.

• BMB Reports
• /
• 제41권5호
• /
• pp.388-393
• /
• 2008

The Pphsc70 (heat shock cognate 70) gene was isolated from the endoparasitoid Pteromalus puparum and then characterized. The full-length cDNA was 2204 base pair (bp) and contained a single 1968 bp ORF that encoded a polypeptide of 656 amino acids with a predicted molecular mass of 71.28 kDa. Phylogenetic analysis based on Hsc70 amino acid sequences from fifteen insect species agreed with the present phylogeny. In addition, genomic DNA confirmed the presence of three introns located at the coding region as well as the 5'UTR. A significant elevation of Pphsc70 expression was observed following heat treatment, however, continued exposure to heat shock or recovery caused the expression of induced mRNA to gradually decline to levels that were significantly lower than those of control pupae (P < 0.05). In addition, a significant increase was observed in the emergence rate of pupae that were preheated at $40^{\circ}C$ and then exposed to $50^{\circ}C$ for 1 h when compared with the pupae that were not preheated, but instead directly exposed to $50^{\circ}C$. Taken together, these results revealed that exposure to gradually increasing temperatures can enhance an insects thermo-tolerance.

• International Journal of Oral Biology
• /
• 제37권3호
• /
• pp.146-152
• /
• 2012

MicroRNAs (miRNAs, miRs) are about 21-25 nucleotides in length and regulate mRNA translation by base pairing to partially complementary sites, predominantly in the 3'-untranslated region (3'-UTR) of the target mRNA. In this study, the expression profile of miRNAs was compared and analyzed for the establishment of miRNA-related odontoblast differentiation using MDPC-23 cells derived from mouse dental papilla cells. To determine the expression profile of miRNAs during the differentiation of MDPC-23 cells, we employed miRNA microarray analysis, quantitative real-time PCR (qRT-PCR) and Alizaline red-S staining. In the miRNA microarray analysis, 11 miRNAs were found to be up- or down-regulated more than 3-fold between day 0 (control) and day 5 of MDPC-23 cell differentiation among the 1,769 miRNAs examined. In qRT-PCR analysis, the expression levels of two of these molecules, miR-194 and miR-126, were increased and decreased in the control MDPC-23 cells compared with the MDPC-23 cells at day 5 of differentiation, respectively. Importantly, the overexpression of miR-194 significantly accelerated mineralization compared with the control cultures during the differentiation of MDPC-23 cells. These results suggest that the miR-194 augments MDPC-23 cell differentiation, and potently accelerates the mineralization process. Moreover, these in vitro results show that different miRNAs are deregulated during the differentiation of MDPC-23 cells, suggesting the involvement of these genes in the differentiation and mineralization of odontoblasts.

• BMB Reports
• /
• 제53권5호
• /
• pp.278-283
• /
• 2020

Muscle fibers are generally formed as multinucleated fibers that are differentiated from myoblasts. Several reports have identified transcription factors and proteins involved in the process of muscle differentiation, but the roles of microRNAs (miRNAs) in myogenesis remain unclear. Here, comparative analysis of the miRNA expression profiles in mouse myoblasts and gastrocnemius (GA) muscle uncovered miR-3074-3p as a novel miRNA showing markedly reduced expression in fully differentiated adult skeletal muscle. Interestingly, elevating miR-3074-3p promoted myogenesis in C2C12 cells, primary myoblasts, and HSMMs, resulting in increased mRNA expression of myogenic makers such as Myog and MyHC. Using a target prediction program, we identified Caveolin-1 (Cav1) as a target mRNA of miR-3074-3p and verified that miR-3074-3p directly interacts with the 3' untranslated region (UTR) of Cav1 mRNA. Consistent with the findings in miR-3074-3p-overexpressing myoblasts, knockdown of Cav1 promoted myogenesis in C2C12 cells and HSMMs. Taken together, our results suggest that miR-3074-3p acts a positive regulator of myogenic differentiation by targeting Cav1.