• Fisheries and Aquatic Sciences
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• 제16권4호
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• pp.291-301
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• 2013

The genomic organization, tissue distribution, and developmental expression of two paralogous GAPDH isoforms were characterized in the mud loach Misgurnus mizolepis (Cypriniformes). The mud loach gapdh isoform genes (mlgapdh-1 and mlgapdh-2) had different exon-intron organizations: 12 exons in mlgapdh-1 (spanning to 4.88 kb) and 11 in mlgapdh-2 (11.78 kb), including a non-translated exon 1 in each isoform. Southern blot hybridization suggested that the mud loach might possess the two copies of mlgapdh-1 and a single copy of mlgapdh-2. The mlgapdh-1 transcript levels are high in tissues requiring high energy flow, such as skeletal muscle and heart, whereas mlgapdh-2 is expressed abundantly in the brain. Both isoforms are differentially regulated during embryonic and larval development, during which their expression is upregulated with the progress of development. Lipopolysaccharide challenge preferentially induced mlgapdh-2 transcripts in the liver. Therefore, the two isoforms have diversified functionally; mlgapdh-1 is associated more closely with energy metabolism, while mlgapdh-2 is related more to stress/immune responses, in the mud loach.

• 한국임상수의학회지
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• 제19권2호
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• pp.195-198
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• 2002

개에서 자연적으로 발생한 12예의 종양에 대해, 종양 억제 유전자 p53의 변이와의 관계를 확인해 보았다. 종양조직에서 일반적인 방법으로 DNA를 추출하여, PCR 기법으로 p53을 증폭하여 염기서열을 확인한 결과, 개의 유선암종 예에서 exon 8의 codon 285에서 CCT $\longrightarrow$ TCT (proline $\longrightarrow$ serine)로 점변이 된 것이 확인되었다. 또한 악성 비만세포 종양 예에서도 exon 8의 codon 249에서 AGT $\longrightarrow$AGC로 점변이 된 것이 확인되었으나 silent point mutation (serine)으로 판명되었다. 이상의 결과를 토대로 개의 유선암종과 악성 비만세포 종양에서 종양억제 유전자 p53의 변이가 확인되었으며, 이는 종양의 형성과 관련된 p53의 역할이나 종양의 치료 및 예후 판정에 p53 을 활용하는 연구의 초석이 되리라 사료되며, 차후 이 유전자에 대한 광범위한 연구가 지속되어야 하리라 생각된다.

• Journal of Plant Biotechnology
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• 제38권1호
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• pp.94-104
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• 2011

Salt stress is a major environmental factor influencing plant growth and development. To identify salt tolerance determinants, we systematically screened salt sensitive rice mutants by use of the Activator/Dissociation (Ac/Ds) transposon tagging system. In this study, we focused on the salt sensitive mutant line, designated SSM-1. A gene encoding a NAC transcription factor homologue was disrupted by the insertion of a Ds transposon into SSM-1 line. The OsNAC075 gene (EU541472) has 7 exons and encodes a protein (486-aa) containing the NAC domain in its N-terminal region. Sequence comparison showed that the OsNAC075 protein had a strikingly conserved region at the N-terminus, which is considered as the characteristic of the NAC protein family. OsNAC075 protein was orthologous to Arabidopsis thaliana ANAC075. Phylogenetic analysis confirmed OsNAC075 belonged to the OsNAC3 subfamily, which plays an important role in response to stress stimuli. RT-PCR analysis showed that the expression of OsNAC075 gene was rapidly and strongly induced by stresses such as NaCl, ABA and low temperature ($4^{\circ}C$). Our data suggest that OsNAC075 holds promising utility in improving salt tolerance in rice.

• International Journal of Industrial Entomology and Biomaterials
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• 제5권1호
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• pp.93-102
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• 2002

We have cloned and characterized the complete fibroin light chain gene from the silkworm Baekok-Jam, Bombyx mori, a recommended variety in Korea. It consists of seven exons and six introns. It consists of 14,663 nucleotides long with an open reading frame of 786 nucleotides that encodes a protein of 262 amino acid residues with a molecular mass of approximately 26,000 dalton. The amino acid alignment revealed that the Baekok-Jam fibroin light chain had 98.5 % protein sequence identity to J139 strain: differed at four amino acid positions (11, 46, 80 and 123). The Northern hybridization analysis showed that the Baekok-Jam fibroin light chain gene was specifically expressed in the posterior silk gland.

• 한국버섯학회지
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• 제9권2호
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• pp.53-58
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• 2011

In order to confirm the presence of putative glucoamylase gene in Tricholoma matsutake genome, the genomic DNA was prepared from T. matsutake NBRC30773 strain and was used as template to clone the glucoamylases gene (TmGlu1). We obtained the nucleotide sequence of TmGlu1 and its franking region. The coding region (from ATG to stop codon) is 2,186 bp. The locations of exons and introns were determined from the nucleotide sequences of 3'- and 5'-RACE PCR and RT-PCR products. On the other hand, to investigate the relationship between composition of medium and glucoamylase expression, we checked the expression level of glucoamylase gene by realtime reverse transcription PCR and measurement of glucoamylase enzyme activity. It was found that enzyme activity of glucoamylase was very low in different medium. Expression of glucoamylases gene appeared to not be affected by different carbon source.

• Journal of Applied Biological Chemistry
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• 제42권4호
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• pp.169-174
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• 1999

Arabidopsis AHL gene contains 4 exons encoding a putative protein highly homologous to the yeast salt-sensitive enzyme HAL2, a 3'(2'),5'-bisphosphate nucleotidase involving in reductive sulfate assimilation. AHL cDNA complemented yeast met22 (hal2) mutant. AHL fusion protein expressed in E. coli exhibited $Mg^{2+}$-dependent, 3'-phosphoadenosine 5'-phosphate (PAP)-specific phosphatase activity. $Li^+,\;Na^+,\;K^+$ and $Ca^{2+}$ ions inhibit the enzyme activity by competing with $Mg^{2+}$ for the active site of the enzyme. The enzyme activity was also sensitive to ${\mu}M$ concentrations of toxic heavy metal ions such as $Cd^{2+},\;Cu^{2+}$ and $Zn^{2+}$, but was not recovered by addition of more $Mg^{2+}$ ions, suggesting that these ions inactivate the enzyme with a mechanism other than competition with $Mg^{2+}$ ions. Inhibition of the AHL enzyme activity may result in accumulation of PAP, which is highly toxic to the cell. Thus, the AHL enzyme could be one of the intial targets of heavy metal toxicity in plants.

• BMB Reports
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• 제37권2호
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• pp.177-184
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• 2004

The Crustacean hyperglycemic hormone (CHH) has been shown to exist as multiple molecular forms in several crustacean species. In Penaeus monodon, a gene encoding CHH (so-called Pem-CHH1) was recently described. In this study, the molecular structures of two other CHH genes (Pem-CHH2 and Pem-CHH3) are reported. Both the Pem-CHH2 and Pem-CHH3 genes contain three exons that are separated by two introns that are similar to the structure of other genes in the same family. An analysis of the upstream nucleotide sequences of each Pem-CHH gene has identified the putative promoter element (TATA box) and putative binding sites for several transcription factors. The binding sites for CREB, Pit-1, and AP-1 were found upstream of all three Pem-CHH genes. A Southern blot analysis showed that at least one copy of each Pem-CHH gene was located within the same 10 kb genomic DNA fragment. These results suggest that the CHH genes are arranged in a cluster in the genome of P. monodon, and that their expression may be modulated by similar mechanisms.

• Food Science and Biotechnology
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• 제16권1호
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• pp.67-70
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• 2007

Mackerel (Scomber japonicus) often causes severe allergic reactions in sensitive people. Food containing undeclared mackerel may pose a risk to such people. The major allergenic protein in fish such as mackerel, codfish, and Alaska pollack has been found to be parvalbumin. In this study, we developed a polymerase chain reaction (PCR) method to detect mackerel DNA using primers corresponding to the parvalbumin gene. We cloned and sequenced 1.5 kb of parvalbumin gene by PCR using mackerel genomic DNA as a template. Nucleotide sequence analysis of genomic parvalbumin gene, composed of 4 exons and 3 introns, allowed the selection of two pairs of oligonucleotide primers specific for mackerel. These primers successfully enabled PCR amplification of specific regions of genomic parvalbumin DNA from mackerel, but no amplification from 8 other fish samples, surimi, and 6 boiled fish pastes. The sensitivity of this method was sufficient to detect 5 ng of purified mackerel DNA mixed with 50 ng of surimi DNA. This rapid and specific method for the detection of allergenic mackerel would be beneficial in reducing food allergy caused by the ingestion of hidden allergen in processed food.

• Journal of Microbiology
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• 제38권4호
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• pp.230-238
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• 2000

Class III chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class III chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5'flanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at -716 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for past-transnational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P chrysogenum and genus Aspergillus.

• Journal of Interdisciplinary Genomics
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• 제1권1호
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• pp.6-9
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• 2019

In this report, the phenotypes of three patients from two families with mucopolysaccharidosis type II (MPS II) are compared: a novel variant and recombinant variant of IDS gene. The results of urine in patients showed a pronounced increase in glycosaminoglycan excretion with decreased iduronate-2-sulfatase enzyme activity in leukocyte, leading to a diagnosis of MPS II. A patient has a novel variant with 1 bp small insertion, c.1224_1225insC in exon 9, which caused frameshifts with a premature stop codon, and two patients have a recombination variant, c.418+495_1006+1304del, leading to the loss of exons 4, 5, 6, and 7 in genomic DNA, which is relatively common in Korean patients. They had different phenotypes even in the same mutation. The patients have now been enzyme replacement therapy with a significant decrease in glycosaminoglycan excretion. Further study on residual enzyme activity, as well as experience with more cases, may shed light on the relationship between phenotypes in MPS II and gene mutations.