• Parasites, Hosts and Diseases
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• v.31 no.3
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• pp.285-292
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• 1993

C. sinensis total RMh was containing large amount of 185 rRNA but little 285 rRNA. The size of the double-stranded cDNA synthesized from poly $(A)^{+}$ mRNA was 0.4-4.2 kb long with tapering unto 9.5 kb. Degenerated oligonucleotides (as 2 sense and 3 antisense Primers) were designed on the conserved regions of the known tropomyosin amino acid sequences. From one out of the PCR amplifications using total CDNA and matrix of primers, a specific gene product, 580 bp in size, was produced. Upon Southern hybridization of the PCR products with Schistosomn mnnsoni tropomyosin (SMTM) CDNA, only one signal appeared at the band of 580 bp product. This 580 bp product was considered to encode C. sinensis tropomyosin (CSTM) and cloned in pGEM-3Zf(-) for DNA sequencing. CSTM cDNA was 575 bp containing one open reading frame of 191 predicted amino acids, which revealed 86.3% homology with SMTM and 51.1% with rrichostronsylur coeubnlormis tropomyosin. CSTM cDNA obtained will serve as a probe in the studies of molecular cloning of CSTM.

• Korean Journal of Plant Tissue Culture
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• v.24 no.2
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• pp.93-97
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• 1997

For the molecular genetic study of cold tolerance mechanism in plants, a cDNA encoding fatty acid desaturase (fad3), converting linoleic acid (18:2, $\omega$-6) to linolenic acid (18:3, $\omega$-3), was isolated from $\lambda$ZAPII Arabidopsis thaliana cDNA expression library by plaque hybridization using fad3 cDNA probe derived from Brassica napus. A 1.8 kb-EcoRI fragment from a lambda clone showing a strong positive hybridization signal was subcloned into pGEM7 and analyzed for its nucleotide sequence. From deduced amino acid sequences, the fad3 gene was revealed to have an open reading frame(ORF) consisting of 386 amino acids with a molecular mass of 44,075 Da. The fad3 gene was compared to chloroplast $\omega$-3 fatty acid desaturase (fad7) and endoplasmic reticulum Δ12 fatty acid desaturase (fad2) to show 70% and 58% amino acid sequence homology, respectively, Especially, amino acids of internal (82 to 151) and carboxy terminal (276 to 333) regions were highly conserved, implying their requisite role for enzymatic functioning of fatty acid desaturases. IPTG-induced fad3 cDNA expression in E. coli cells was suggested to be toxic to bacterial growth.

• Journal of Veterinary Clinics
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• v.25 no.3
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• pp.139-145
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• 2008

To investigate the genotypic diversity of the porcine reproductive and respiratory syndrome viruses (PRRSV) in Korea, we examined 92 clinical samples from three provinces by RT-PCR and a nested PCR, and the complete open-reading frame 7 (ORF 7) sequences of 15 samples selected from 72 PCR-positive specimens were analyzed. When we compared nucleotide (amino acid) sequences of 80 isolates from Korea and overseas countries, the sequences of 7 samples belonged to North American (NA)-genotype, and those of 8 samples, to European (EU)-genotype. The nucleotide (amino acid) identities between two genotypes were 63.7% (59.8%) to 65.1% (63.1%). When compared with NA prototype VR-2332, the 7 strains of NA-genotype shared 89.8% (93.6%) to 91.2% (96.0%) identity of nucleotide (amino acid) sequence. The 8 strains of EU-type shared 93.6% (92.3%) to 94.3% (93.8%) identity of nucleotide (amino acid) sequence as compared to EU prototype Lelystad. In phylogenetic tree analysis by neighbor-joining method, all of the 8 EU-type strains were clustered into group 4 distinct from ED-prototype Lelystad (group 1). In NA-genotype, 24 domestic isolates reported previously and the 7 strains of NA-type determined in this study were clustered into group 1, while US prototype VR 2332 was classified into different group (group 2). These results suggest that emergence of EU-genotype and the dual-infection of NA- and EU-genotypes may be prevalent in the pig farms in Korea. The high degree of genetic diversity of field PRRSVs should be taken into consideration for control and preventive measures.

• Korean Journal of Microbiology
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• v.41 no.3
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• pp.157-163
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• 2005

The acetate kinase gene from the copepod Paracyclopina nana was cloned. The open reading frame (ORF) was 1,200 bp, and poly(A) signal sequence was located in the end of the ORF. After the molecular phylogenetic analysis of P nana acetate kinase gene, it was revealed that it formed the same branch with that of Aspergillus. Also P. nana acetate kinase showed the difference with those of other prokaryotic microorganisms but showed the same clade with those of fungi. We also confirmed that the recombinant protein of P. nana acetate kinase made approximately 50 kDa after expression of recombinant gene construct in E. coli. This may be useful to compare this protein to those of other organisms in biochemical characteristics.

• The Korean Journal of Mycology
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• v.25 no.1 s.80
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• pp.1-5
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• 1997

The cDNA encoding ${\beta}-tubulin$ of Pleurotus sajor-caju was isolated using an internal gene segment probe amplified by polymerase chain reaction (PCR) of genomic DNA and by cDNA library screening. The cDNA was consisted of 1560 nucleotides(nt), including a 5'-untranslation region (UTR) of 27nt, an open reading frame (ORF) of 1341nt, and a 3'-UTR of 191nt. The ORF encoded a protein of 446 amino acids(aa), which shows over 80% homology with ${\beta}-tubulins$ of other filamentous fungi. Southern hybridization analysis showed that there were two isotypes of ${\beta}-tubulin$ genes in P. sajor-caju. Through sequence analysis we found that ${\beta}-tubulin$ had a unusual $Cys^{165}$ residue, which might be a significant factor for the insensitivity of fungi to fungicide benomyl.

• The Plant Pathology Journal
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• v.27 no.3
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• pp.285-290
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• 2011

Fusarium graminearum virus 2 (FgV2) infects Fusarium graminearum strain 98-8-60 and has at least five segments of double-stranded RNAs (dsRNAs), denoted as dsRNA-1 to dsRNA-5. In this study, the genome of FgV2 was sequenced and its phylogenetic relationship with other mycoviruses was analyzed. The lengths of FgV2 dsRNAs 1-5 ranged from 2414 to 3580 base pairs (bp). The 5' and 3' untranslated regions (UTRs) are highly conserved, and each dsRNA segment had 78-105 and 84-306 bp of 5' and 3' UTRs, respectively. Each dsRNA segment contained a single open reading frame (ORF). Computer analysis of dsRNA-1 revealed a putative open reading frame (ORF) that shows high sequence identity with an RNA-dependent RNA polymerase (RdRp) containing eight conserved motifs. dsRNAs 2-5 also each contain one putative ORF coding for products of unknown function. The sequences of FgV2 dsRNA-2 and dsRNA-3 have significant sequence identity with Magnaporthe oryzae chrysovirus 1 (MoCV1) dsRNA-3 and -4, respectively. When compared to other dsRNA mycoviruses in a phylogenetic analysis of the putative RdRp protein, FgV2 was found to form a distinct virus clade with Aspergillus mycovirus 1816 and MoCV1 in the family Chrysoviridae.

• Microbiology and Biotechnology Letters
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• v.28 no.3
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• pp.147-151
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• 2000

School of Food Biotechnology, W0050ng University, San 7-6, Jayang~dong. Dong-ku1 Taejon 300-100, Korea - The nucleotide sequence of approximately 2.4 kb immediately adjacent to ptsG gene coding for the glucose permease of Brevibacterium ammoniagenes was detennined. A putative open reading frame (ORP) of 1.467 nucleotides encoding a polypeptide of 489 amino acid residues and a TAA stop codon was identified. The deduced amino acid sequence of the ORF product has a high homology with the 30S ribosomal protein S 1 of Mycohacteriwn tuberculosis (83 % ). M leprae (74%), Streptomyces coelicola (77%), and Escherichia coli (40%). suggesting that the predicted product of ORF is a ribosomal protein S 1. The ORF is located at a distance of 266 nucleotides upstream from ptsC gene with a same translational direction.

• Korean Journal of Plant Resources
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• v.24 no.5
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• pp.584-590
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• 2011

Wound inducible P450-Esg cDNA, one of cytochrome P450 gene family, was isolated from shoot of Euiseong garlic cultivar. P450-Esg cDNA possesses highly conserved heme-binding domain in the nucleotide sequence, and 1,419 bp of open reading frame (ORF) coding of 473 amino acids. Based on the nucleotide sequence analysis of P450-Esg homologous from twelve garlic cultivars, two domains, one domain between 472 to 510 bp, and the other between 1,210 to 1,249 bp from start codon (ATG), showed various nucleotide polymorphism among cultivars. Sequence of heme-binding domain in P450-Esg homologous, which is located at the domain between 1,210 to 1,240 bp from start codon, showed various nucleotide polymorphism as well as amino acid sequence polymorphism among twelve garlic cultivars. Anther domain, between 472 to 510 bp from start codon, showed exactly same amino acid sequence in the twelve garlic cultivars, but there were various single nucleotide polymorphism to the cultivars.

• Korean Journal of Microbiology
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• v.39 no.2
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• pp.83-88
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• 2003

We cloned a gene encoding acetyl xylan esterase(axeA) of Streptomyces coelicolor A3(2) and studied its expression pattern in Escherichia coli. The full sequence of axeA was amplified by PCR. Sequence analysis of the PCR product revealed an open reading frame of 1,008 nucleotides encoding a protein consisted of 335 amino acid residues, with a calculated molecular mass of about 38 kDa. The base sequence showed 98% homology to the same gene of Streptomyces lividans. Two different kinds of acetyl xylan esterases were produced in Escherichia coli(pLacI) by IPTG induction; their molecular weights were 38 kDa and 34 kDa, respectively. Of these, 38 kDa protein seemed to be a total protein holding N-terminal signal peptide region, whereas 34 kDa protein seemed to be a matured protein without signal peptide which was produced by peptide bond cleavage between two amino acid residues of alanine 41 and alanine 42.

• Korean Journal of Microbiology
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• v.42 no.1
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• pp.67-72
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• 2006

The nucleotide sequence of the cloned cellulolytic xylanase gene (bglBC2) from B. circulans ATCC21367 was determined. bglBC2 consists of an 1,224 bp open reading frame (ORF) coding for a polypeptide of 407 amino acids with a deduced molecular weight of 45 kDa. The Shine-Dalgarno (SD) sequence (5'-AAAGGAG-3') was found 9 bp upstream of the initiation codon, ATG. A promoter region corresponding closely to the B. subtilis consensus sequence (-35: TTGACA,-10: TATAAT) was detected, the putative -35 and -10 sequences of which were TTTACA and TATACT, respectively. The deduced amino acid sequence of the cellulolytic xylanase showed 97% homology with that of the alkaline $endo-\beta-1,4-glucanase$ from B. circulans KSM-N257, 75% homology with that of the $endo-\beta-1,3-1,4-glucanase$ from B. circulans WL-12, and 45% homology with that of the $endo-\beta-1,4-glucanase$ (cellulase) from Bacillus sp. KSM-330. The bglBC2 sequence was deposited in Gen-Bank under the accession number AY269256.