• Biomedical Science Letters
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• v.23 no.4
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• pp.416-420
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• 2017

Endoplasmic reticulum (ER) stress induces unfolded protein response (UPR) via inositol-requiring enzyme 1 (IRE1) activation, which sends a molecular signal for X box-binding protein 1 (XBP1) mRNA splicing in the cytosol. IRE1 endoribonuclease activity induces cleavage of XBP1 mRNA. The XBP1 mRNA is then ligated by an uncharacterized RNA ligase and translated to produce spliced XBP1 by 23 nt removed in which contains the PstI restriction enzyme site. The splicing of XBP1 mRNA can be detected by semiquantitative RT-PCR, and then splicing of XBP1 is a useful tool to measure the genetic variability in ER stress. In this study, we have estimated IRE1-dependent splicing of XBP1 mRNA under conditions of various hypothermia. The results indicated that hypothermia regulated ER stress. This study demonstrated that hypothermia is closely related to ER stress and may be useful for early diagnosis of ER-associated disease.

• Journal of Microbiology and Biotechnology
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• v.10 no.4
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• pp.559-563
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• 2000

A dextransucrase gene (dsrB742) that expresses a dextransucrase to synthesize mostly ${\alpha}-1{\rightarrow}6$ linked dextran with a low amount (3-5%) of ${\alpha}-1{\rightarrow}3$ branching was cloned and sequenced from Leuconostoc mesenteroides B-742CB. The 6.1-kb PstI fragments were ligated with pGEM-3Zf(-) and transformed into E. coli $DH5{\alpha}$. The recombinant clone (pDSRB742) synthesized dextran on an agar plate containing 2% (w/v) sucrose. The dextran synthesized was hydrolyzed with Penicillium endo-dextranase. The hydrolyzate was composed of glucose, isomaltose, isomaltotriose, and branced pentasaccharide. The nucleotide sequence of dsrB742 showed one open reading frame (ORF) composed of 4,524 bp encoding dextrasnsucrase. The deduced amino acid sequence revealed a calculated molecular mass of 168.6 kDa. It also showed an activity band of 184 kKa on a non-denaturing SDS-PAGE (10%). The amino acid sequence of DSRB742 exhibited a 50% similarity with DSRA from L. mesenteroides B-1299, a 70% similarity with DSRS from L. mesenteroides B-512 (F, FMCM) and a 45-56% similarity with Streptococcal GTFs.

• The Journal of the Korea Contents Association
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• v.6 no.10
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• pp.114-125
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• 2006

In the previous work, we presented HDT for locating logical errors in Java programs. The HDT locates an erroneous method at an execution tree using an algorithmic program debugging technique and locates a statement with errors in the erroneous method using a step-wise program debugging. It reduced the number of programmer debugging in Java programs. But the HDT still increases the number of debugging because the size of the recent programs increases than the past programs and the number of methods is increasing. This paper proposes HDTS using a program slicing technique (PST) at the MDT. HDTS can reduce the number of programmer debugging. Specially, the more the number of methods and statements increases, the more HDTS has effects.

• Journal of Microbiology and Biotechnology
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• v.1 no.4
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• pp.240-245
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• 1991

We have constructed a promoter-probe vector pKU20 using pKT230, a derivative of broad-host-range plsmid RSF1010, as a base. The pKU20 contains structural gene for aminoglycoside phos-photransferase (aph), without promoter, and a multiple cloning site upstream the aph. Using this vector, a 412base pairs (bp) PstI fragment showing strong promoter activity both in Escherichia coli LE392 and Pseudomonas putida KCTC1644 has been cloned from Pseudomonas fluorescens chromosomal DNA on the basis of streptomycin resistance. The nucleotide sequence of the 412 bp fragment has been determined and the putative - 35 and -10 region was observed. Insecticidal protein gene of Bacillus thuringiensis subsp. kurstaki HD-73 inserted on downstream of the promoterlike DNA fragment was efficiently expressed in E. coli and P. putida. The toxin protein was efficiently synthesized in an insoluble form in both strains.

• Korean Journal of Microbiology
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• v.30 no.6
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• pp.539-545
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• 1992

Recombinant plasmid pPLac15 determined both phosphoenolpyruvate-dependent phosphotransferase uptake of lactose and phospho-$\beta$-galactosidase (Moon et al., 1989). A restriction mapping of the pPLac15 was compiled with several restriction enzymes and a seriese of sub clones into pUC18 was constructed. From an analysis of the proteins produced by Escherichia coli cells of transformants containing each of the recombinant subclone plasmids, it was found that the gene for phospho-$\beta$-galactosidase in pUCI8 was expressed about 1.8-folds in E. coli.

• Journal of Environmental Science International
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• v.20 no.4
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• pp.551-554
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• 2011

The production of the sharp-toothed eel by commercial catch off waters of Korea is annually declined after 1978. This study was carried out to obtain the stock management of the sharp-toothed eel using the PCR-aided RFLP method. The mtDNA COI gene was amplified using species-specific primers and PCR product was observed to 700 bp. Amplified DNA fragments were treated with six kinds of restriction enzymes (BaeHI, EcoRI, PstI, Ksp22, HinfI and HaeIII). The treatment of HaeIII showed a distinct PCR product between Yeosu/Jinhae/Jeju/Goseoung and Jangheung/Haenam populations that were observed from 300 to 400 bp in reference to 100 bp molecular marker. However, DNA fragment within populations had an identical pattern. The phylogenetic homology is 82% between two populations inferred from RFLP PCR product pattern using NTsysPC ver. 2.1. The use of HaeIII plays an important role in discriminating populations. It is thought that adults after over-wintering in the southern part of Jeju migrate to the Yeosu, Jinhae and Goseoung regions to spawn instead of to southwestern waters. Individuals within populations showed a relatively active genetic mixing and migration regardless of geography. However, the genetic ancestor of Jangheung and Haenam populations is appeared to be more adjacent to China or Japan than Jeju.

• Korean Journal of Agricultural Science
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• v.23 no.1
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• pp.99-107
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• 1996

Eighteen nuclear probes were used to examine RFLP(restriction fragment length polymorphism) between six species of Artemisia spp. of Korea. Total DNA from six different species of Artemisia was separately cut with three restrict enzymes. The PstI enzyme was showed to reduce the variation of polymorphisms than the other two enzymes(EcoRl and BamHI). The genetic variation of polymorphism was similar between the Dhewegiki-ssug and Cham-ssug. RAPD analysis was applied to the same six species of Artemisia spp. in order to assess the degree of DNA polymorphism within the Artemisia genus. Six species of Artemisia were evaluated for variation using a set of 11 random 10-mer primers. Nine out of the eleven primers revealed scorable polymorphisms between six species of Artemisia spp. Genetic distances between each of the species were calculated and cluster analysis was used to generate a dendrogram showing phylogenetic relationships between them This result indicates that molecular markers will be more usable in intraspecific study of Artemisia spp. than isoenzyme markers.

• The Journal of the Korean Society for Microbiology
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• v.35 no.1
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• pp.49-60
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• 2000

ITSI-5.8S-ITSII rDNA region was amplified from the reference strains and clinical isolates with ITS1 and ITS4 primers. These primers amplified DNA fragments of 550 bp in Microsporum audouinii and Trichophyton violaceum, 700 bp in Microsporum gypseum, Trichophyton mentagrophytes, Trichophyton rubrum, and Trichophyton tonsurans, and 750 bp in Microsporum ferreugineum and Microsporum canis. The restriction enzyme patterns of PCR products digested with 13 restriction enzyme including PstI were distint among the genera, whereas identical in the same species. Examination of the ITS (Internal Transcribed Spacers)1 nucleotide sequence revealed that there was the genetic difference in each genera and species. Phylogenetic relationship among each species showed that the Trichophyton mentagrophytes was more closely related Trichophyton tonsurans than Trichophyton rubrum, and Microsporum gypseum was less related than Microsporum spp..

• Clinical and Experimental Pediatrics
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• v.45 no.12
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• pp.1585-1590
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• 2002

Rhizomelic chondrodysplasia punctata(RCDP) is a rare autosomal recessive disorder clinically characterized by symmetrical shortening of the proximal limbs, contractures of joints, a typical dysmorphic face, cataracts, and itchyosis. Patients with RCDP can be subdivided into three subgroups based on biochemical analysis and complementation studies. RCDP type I results from mutations in the PEX7 gene encoding the peroxisomal targeting signal type II(PST2) receptors and presents with both a defect in plasmalogen biosynthesis and phytanic acid oxidation. RCDP type II is deficient in the activity of dihydroxyacetonephosphate acyltransferase(DHAP-AT). RCDP type III is deficient in alkyl-dihydroxyacetonephosphate synthase(alkyl-DHAP). We report a case of RCDP type I which was confirmed with biochemical study, fibroblast culture, and gene study.

• Journal of Microbiology and Biotechnology
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• v.12 no.5
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• pp.716-721
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• 2002

The ldhL gene encoding the $_L$-(+) lactate dehydrogenase was cloned from Lactobacillus reuteri ATCC 55739 chromosomal DNA and characterized. An internal 750-bp fiagment of ldhL gene was amplified by PCR using primers based on the conserved region of lactobacilli ldhL genes. A genomic library off. reuteri ATCC 55739 was constructed using pBR322, and colony hybridization experiments were performed using the 750-bp fragment as aprobe. One clone harboring a 4.0-kb PstI fragment was identified, and nucleotide sequencing confirmed it as an open reading frame of 972 bp in size in the middle. In addition to IdhL gene, an ORF homologous to Streptococcus pneumoniae TIGR4 hydrolase gene and 3' part of phosphomevalonate kinase gene (mvaK2) were also found on the 4 kb fragment. $_L$-LDH of L. reuteri ATCC 55739 showed the highest degree of homology with the $_L$-LDH of Pediococcus acidilactici (62.4%), fullowed by the $_L$-LDH of Lactobacillus pentosus (58.7%). The size of IdhL transcript determined by Northern blot was 1 kb, indicating the monocistronic nature of IdhL.