• Journal of Mushroom
• /
• v.2 no.3
• /
• pp.145-148
• /
• 2004

This study was carried out to screen the fruiting body formation-specific genes from the medicinal mushroom Cordyceps militaris. A cDNA synthesized using total RNA from 4 stages of mushroom development, mycelium, primordium, immature fruiting body and mature fruiting body. Differential expression gene screening was performed by DD-PCR(Differential Display Arbitrary Primer PCR) with cDNA, we sequenced partial 6 genes using pGEM cloning vector. The DNA Sequence of the six DD-PCR products derived from differentially expressed genes was compared to that in the GenBank database by using the NCBI BLAST search to identify similarities to known sequences. Sequence analysis showed that six of DD-PCR products have unknown sequence.

• Archives of Pharmacal Research
• /
• v.28 no.2
• /
• pp.232-237
• /
• 2005

Mast cells play an important role in allergic inflammation by releasing their bioactive mediators. The function of mast cells is enhanced by stimulation because of the induction of specific genes and their products. While many inducible genes have been elucidated, we speculated that a significant number of genes remain to be identified. Thus, we applied differential display (dd) PCR to establish a profile of the induced genes in bone marrow-derived mast cells (BMMCs) after they were co-cultured with 3T3 fibroblasts. To date, 150 cDNA fragments from the connective-type mast cells (CTMCs) were amplified. Among them, thirty cDNA fragments were reamplified for cloning and sequencing. The ddPCR strategy revealed that serine proteases were the most abundant genes among the sequenced clones induced during the maturation. Additionally, unknown genes from the co-culture of BMMCs with 3T3 fibroblasts were identified. We confirmed their induction in the CTMCs by Northern blot analysis and RT-PCR. Characterization of these induced genes during the maturation processes will provide insight into the functions of mast cells.

• BMB Reports
• /
• v.33 no.2
• /
• pp.184-187
• /
• 2000

Prostaglandin $E_2$ ($PGE_2$) plays an important role in the regulation of various gastric functions, and the growth-inhibitory activities on tumor cells are studied in vitro and in vivo. Although the mechanisms have attracted many researchers in the past decade, the molecular mechanisms of cell cycle arrest, or induction of apoptosis by $PGE_2$, is unclear. We investigated the effects of $PGE_2$ on the growth of the human gastric carcinoma cell line SNU1 and genes that are regulated by $PGE_2$ and isolated them using differential display RT-PCR (DD RT-PCR). FACS analysis suggested that SNU1 cells were arrested at the G1 phase by $PGE_2$ treatment. This growth inhibitory effect was in a time- and dose-dependent manner. Treatment of SNU1 cells with $10\;{\mu}g/ml$ $PGE_2$, followed by DD RT-PCR analysis, revealed differently expressed bands patterns from the control. Among the differently expressed clones, we found an unidentified cDNA clone (HGP-27) overexpressed in $PGE_2$-treated cells. The full-length cDNA of HGP-27 was isolated using RACE, which consisted of a 30-nt 5'-noncoding region, a 891-nt ORF encoding the 296 amino acid protein, and a 738-nt 3'-noncoding region including a poly(a) signal. This gene was localized on the short arm of chromosome number 11. Using the Motif Finder program, a myb-DNA binding repeat signature was detected on the ORF region. The COOH-terminal half was shown to have similarity with the $NH_3$-terminal domain of thioredoxin (Trx). This relation between HGP-27 and Trx implied a potential role for HGP-27 in modulating the DNA binding function of a transcription factor, myb.

• Molecular & Cellular Toxicology
• /
• v.2 no.4
• /
• pp.251-256
• /
• 2006

The exploration of genes which expressions are changed by exposure to ecotoxicants or pollutants can provide the important information about the reaction mechanisms in the body as well as adaptation to exterior stimulus or environmental changes. Also they can be developed as biomarkers for the detection of environmental pollution. Differential display polymerase chain reaction (DD-PCR) technique has been usefully used to hunt the clones which expressions are up-regulated or down-regulated by exterior changes and this study aimed to search for those clones in diesel oil-exposed rockfish (Sebastes schlegeli) using DD-PCR. The RNA isolated from liver of 20 ppb diesel oil-exposed rockfish was used for screening of the differentially displayed genes and total 44 differentially expressed genes (DEG) are detected then their nucleotide sequences were analyzed. The present data provided the general information about the effect of diesel oil contamination on marine organism and further more the primary step in development of new biomarkers for marine environmental pollution or ecotoxicological stresses.

• Korean Journal of Breeding Science
• /
• v.42 no.5
• /
• pp.565-573
• /
• 2010

Thinning of apple fruitlets is one of the most laborious and important works for the improvement of fruit quality and for the promotion of sufficient flower bud formation to prevent alternate bearing in commercial cultivars. Lateral fruits of self-thinning apple cultivars fall naturally within 30 days after full bloom and only central fruit remains to mature. Differences of gene expression between central fruit and lateral fruit were investigated by differential display (DD) PCR. Partial cDNAs of 30 clones from the central fruit and 24 clones from the lateral fruit were selected for nucleotide sequence determination and homology searches. The levels of transcripts coding for proteins involved in pathogenesis related proteins, senescence, temperature stress, protein degradation, fruit browning, sorbitol metabolism were significantly higher in pedicels of lateral fruit than in pedicels of central fruit. On the other hand, the up-regulation of proteins involved in anthocyanin and flavanol biosynthesis and ethylene synthesis were observed in pedicels of central fruit. In Real time PCR analysis, cytochrome P450 gene was confirmed as showing a higher expression level in lateral fruit than in central fruit. The results of this study indicate that differentially expressed genes are related to self-thinning characteristics in apple tree.

• Mycobiology
• /
• v.30 no.2
• /
• pp.88-92
• /
• 2002

Rehmannia glutinosa is a perennial medicinal plant belonging to the family Scrophulariaceae with more than 300 species known in the world, especially in temperate regions. Its roots have been used widely in Korea for medicinal purposes. However, it is commonly infected by various pathogens during storage, causing great damage to the roots, and impedes the intensive farming of the crop. Therefore, an attempt has been made to isolate and screen a resistance gene against the pathogen Fusarium oxysporum using differential display. We treated salicylic acid(SA), and isolated a resistance gene that responds to SA. As a result, we found that SA was involved in plant defense mechanism in pathogenicity tests with SA treated and non-treted plants, and we isolated a partial PR-la gene through differential display polymerase chain reaction(DD-PCR) method.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
• /
• 2005.05a
• /
• pp.239-242
• /
• 2005

본 연구는 성장 속도 및 서로 다른 육질 특성을 지닌 돼지 품종을 이용하여, 육질 및 성장에 관련된 유전자원을 확보하고, 이를 이용한 유전 육종의 기초 자료를 제공하기 위하여 수행하였다. Differential display (DD) RT-PCR 기법을 통해 돼지 품종 간 발현 차이를 보이는 유전자인 NADH dehydrogenase 1과 ATPase 6를 동정하였다. 동정된 유전자의 발현량 분석을 위한 RT-PCR 결과, 각 유전자의 발현량이 재래돼지에서 외래 품종 (랜드레이 스 및 요크셔)에 비해 2배 이상 높음을 확인 할 수 있었다 (p<0.01). 이러한 발현차이 유전자를 이용하여 육질과의 관련성 연구 및 유전자의 기능에 대한 연구가 지속되어야 할 것이다.

• International Journal of Oral Biology
• /
• v.37 no.4
• /
• pp.167-173
• /
• 2012

This study investigated the genes involved in the differentiation of odontoblasts derived from human dental pulp stem cells (hDPSCs). hDPSCs isolated from human tooth pulp were validated by fluorescence activated cell sorting (FACS). After odontogenic induction, hDPSCs were analyzed investigated by Alizaline red-S staining, ALP assay, ALP staining and RT-PCR. Differential display-polymerase chain reaction (DD-PCR) was performed to screen differentially expressed genes involved in the differentiation of hDPSCs. By FACS analysis, the stem cell markers CD24 and CD44 were found to be highly expressed in hDPSCs. When hDPSCs were treated with agents such as ${\beta}$-glycerophosphate (${\beta}$-GP) and ascorbic acid (AA), nodule formation was exhibited within six weeks. The ALP activity of hDPSCs was found to elevate over time, with a detectable up-regulation at 14 days after odontogenic induction. RT-PCR analysis revealed that dentin sialophosphoprotein (DSPP) and osteocalcin (OC) expression had increased in a time-dependent manner in the induction culture. Through the use of DD-PCR, several genes were differentially detected following the odontogenic induction. These results suggest that these genes may possibly be linked to a variety of cellular process during odontogenesis. Furthermore, the characterization of these regulated genes during odontogenic induction will likely provide valuable new insights into the functions of odontoblasts.

• Parasites, Hosts and Diseases
• /
• v.37 no.4
• /
• pp.257-263
• /
• 1999

Identification of the genes responsible for the recovery of virulence in brain-passaged Acanthamoeba culbertsoni was attempted via mRNA differential display polymerase chain reaction (mRNA DD-PCR) analysis. In order to identify the regulatory changes in transcription of the virulence related genes by the brain passages, mRNA DD-PCR was performed which enabled the display of differentially transcribed mRNAs after the brain passages. Through mRNA DD-PCR analysis. 96 brain-passaged amoeba specific amplicons were observed and were screened to identify the amplicons that failed to amplify in the non-brain-passaged amoeba mRNAs. Out of the 96 brain-passaged amoeba specific amplicons, 12 turned out to be amplified only from the brain-passaged amoeba mRNAs by DNA slot blot hybridization. The clone, A289C, amplified with an arbitrary primer of UBC ＃289 and the oligo dT$_{11}$-C primer, revealed the highest homology (49.8％) to the amino acid sequences of UPD-galactose lipid transferase of Erwinia amylovora, which is known to act as an important virulence factor. The deduced amino acid sequences of an insert DNA in clone A289C were also revealed to be similar to cpsD, which is the essential gene for the expression of type III capsule in group B streptococcus. Upregulated expression of clone A289C was verified by RNA slot blot hybridization. Similar hydrophobicity values were also observed between A289C (at residues 47-66) and the AmsG gene of E. amylovora (at residues 286-305: transmembrane domains). This result suggested that the insert of clone A289C might play the same function as galactosyl transferase controlled by the AmsG gene in E. amylovora.a.

• The Journal of Natural Sciences
• /
• v.11 no.1
• /
• pp.95-98
• /
• 1999

Chinese cabbage (Brassica campestris), one of the major vegetable crops in Korea, undergoes self-incompatible pathway for reproduction. To maintain inbred lines of chinese cabbage, a method in that $CO_2$ gas is treated to the pistils to break the self-incompatibility and thereby self-pollens can successfully make germination and fertilization has been selectively used in speed company. In this study, the pistil genes induced by the $CO_2$ treatment was investigated by mRNA differential display (DD-PCR) method. The result shows PCR products amplified in a differential pattern from both $CO_2$ gas treated- and untreated-pistil mRNAs, suggesting that the pistil genes are probably regulated positively and also negatively by the $CO_2$ gas.