• Korean Journal of Microbiology
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• v.40 no.1
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• pp.12-16
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• 2004

Acid proteinase has been discovered in Aspergillus niger (acid protease A) and Cryphonectria parasitica (acid proteinase EapC) and it plays major roles in cheese formation from milk. In this study, a partial gene encoding acid proteinase in Penicillium oxalicum HCLF-34 was cloned by using PCR with degenerate primers corresponding to highly conserved regions of the acid proteinase. The partial acid proteinase gene in P. oxalicum HCLF-34 contains an open reading frame of 438 base pairs and encodes an acid proteinase protein of 146 amino aicds. The predicted amino acid sequences showed 71 % homology with acid protease A and 67% homology with EapC.

• Korean Journal of Microbiology
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• v.35 no.1
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• pp.25-27
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• 1999

Degenerate primers corresponding to the sequences of the copper-binding regions in the fungal laccases were used to isolatc laccase gene specific fragment by PCR in Coprinus congregahts. A 144 bp DNA hagrnent was cloned and was identified to have 60-69 % homology with other fungal laccase genes. The predicted amino acid sequcnces showed 68-75% homology with other fungal laccase proteins.

• Korean Journal Plant Pathology
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• v.11 no.1
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• pp.73-79
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• 1995

Complementary DNAs (cDNAs) to the coat protein gene of an ordinary Korean strain of potato virus Y (PVY-OK) isolated from potato (cv. Superior) were synthesized and cloned into a plasmid pUC119 and sequenced. The RNA of the virus propagated in tobacco (Nicotinaa sylvestris) was extracted by the method of phenol extraction. The first strand of cDNAs to the coat protein penomic RNA of the virus was made by Moloney murine leukemia virus reverse transcriptase. The cDNA were synthesized and amplified by the method of polymerase chain reaction (PCR) using a pair of oligonucleotide primers. PVYCP3P and PVYCP3M. The size of cDNAs inserted in pUC119 plasmid was estimated as about 840 bp upon agarose gel electrophoresis. Double stranded cDNAs were transformed into the competent cell of E. coli JM109. Sequence analysis of cDNAs was conducted by the dideoxynucleotide chain termination method. Homology of cDNAs of the PVY-OK coat protein genomic RNA with those of PVY-O (Japan), PVY-T (Japan), PVY-TH (Japan), PVYN (The Netherlands),and PVYY (France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. Homology at the amino acid level turned out to the be 97.4%, 92.5%, 92.9%, 92.9% and 98.5%, respectively.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.39-39
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• 2003

Methylation of DNA 5-cytosine in mammalian early embryo affects great deal in nuclear reprogramming and chromatin remodeling of developing embryo. Current efforts to clone and produce cloned animals including transgenic animals face various problems including low birth rate, irregular development, and so on. In this report, cDNA for the one of house keeping methyltransfcrase, Dnmt1 was cloned from bovine somatic tissues and was analyzed for its nucleotide sequences to investigate the structure and function of the gene in bovine early development. Nucleotide sequence of bovine Dnmt1 homologue showed 76.8% identity with that of human Dnmtl and 66.4% with mouse Dnmt1. Translated amino acid sequence showed 88.4% homology with human homologue and 75.8% homology with mouse counterpart. Three types of Dnmt1 are reported in mouse and human, and are likely present in bovine tissues. Understanding of role of Dnmt1 in bovine development may shed a light in the field of animal, especially bovine cloning.

• Korean Journal of Microbiology
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• v.30 no.6
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• pp.507-513
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• 1992

Serratia marecescens RH1 isolated from soil samples produced large amount of extracellular proteases. One of the genes encoding an extracellular protease form S. marcescens RH1 was cloned in Escherichia coli by shot gun cloning method. The cloned protease, SSP, was stably expressed by its own promoter and excreted into the extracellular medium from E. coli host (ORF) of 3.135 nucleotides corresponding to 1.045 amino acids (112 kDa). The nucleotide and deduced amino acid sequence of SSP showed high overall homology (88%) to one of the S. marcescens protease (27), but low homology to other serine protease families. The optimal pH and temperature of the enzyme were pH 9.0 and 45.deg.C respectively. The activity of protease was inhibited by phenylmethylsulfonyl fluoride (PMSF), which suggests that the enzyme is a serine protease.

• Korean Journal of Microbiology
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• v.32 no.2
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• pp.115-119
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• 1994

Replication control region of pSBK203, a chloramphenicol acetyltransferase conferring plasmid from Staphylococus aureus was cloned and its nucleotide sequence has been determined. Base sequence homology of this copy control region with those of plasmids belonging to pT181 family was obtained and analyzed. Copy number of four copy mutants derived by addtion or deletion of nucleotides in unique XbaI recognition site in copy control region of pSBK203 was also determined.

• Journal of the Korean Magnetic Resonance Society
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• v.14 no.2
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• pp.105-116
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• 2010

SH3YL1, a novel protein containing one Src homology 3 domain at the carboxyl terminus was first detected in mouse anagen skin cDNA. This protein had a significant homology with YHRO 16c/Ysc 84, the yeast Src homology 3 domain-containing protein. The sequence identity was remarkable at the carboxyl and amino-terminal Src homology 3 domain, suggesting that the novel protein is a mouse homolog of the yeast protein and thus was termed as SH3YL1. SH3YL1 is composed of two domains, a DUF500 at N-termini and a SH3 domain at C-termini. In our study we cloned the SH3 domain in bacterial expression system in Escherichia coli using pET32a vector with TEV protease cleavage site and purified as a monomer using affinity chromatography. The N-terminal poly-Histidine tag was cleaved with TEV protease and target protein was used for backbone studies. Our study showed that SH3 domain primarily consists of $\beta$-sheet which is in consistence with previous result performed on the truncated SH3 domain of SH3YL1.

• Applied Biological Chemistry
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• v.40 no.1
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• pp.30-33
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• 1997

About 250 bp ura5 gene (Orotate phosphoribosyl transferase) fragment was cloned from genomic DNA of entomopathogenic fungus Metarhizium anisopliae by using PCR method. Entire nucleotide sequences of cloned DNA fragment were determined and analysed as compared with other fungus ura5 genes. The amino acid sequence deduced from the nucleotide sequence showed 85.5% homology to ura5 protein of Trichoderma reesei. Using this 250 bp PCR fragment we have isolated full ura5 gene of M. anisopliae by genomic Southern hybridization and the isolated 4.4 kb DNA fragments were mapped by restrictional enzyme.

• International Journal of Industrial Entomology and Biomaterials
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• v.4 no.1
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• pp.63-68
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• 2002

A cDNA encoding a putative member of cathepsin B of the thiol pretense superfamily was cloned from a cDNA library of the mulberry longicorn beetle, Apriona germari. Sequence analysis of the cDNA encoding the cathepsin B of A. germari (AgCatB) revealed that the 972 bp cDNA has an open reading frame of 324 amino acid residues. The deduced protein sequence of the AgCatB showed high homology with cathepsin B of the insects, Bombyx mori (47.3% amino acid identity), Helicoverpa armigera (46.6%) and Sarcophaga peregrina (45.6%), and the lowest homology with Aedes aegypti (33.2%). The AgCatB contains six disulfate bonds typical for cysteine pretenses. The three amino acid positions Cys-109, His-267, and Asn-287 which are conserved, active sites characteristic for cathepsin B, were also found. Phylogenetic analysis further confirmed that the AgCatB has a close relationship with that of B. mori, H. armigera and S. peregrina.

• The Journal of Korean Society of Virology
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• v.30 no.1
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• pp.1-10
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• 2000

The nonstructural protein NSP4, encoded by gene 10 of rotavirus, has been shown to playa role in viral assembly and known to be an enterotoxin, causing diarrhea in mouse pups. NSP4 gene was cloned from CBNU-2 (virulent bovine rotavirus/diarrheic fecal sample) and CBNU-1 (cell-culture adapted bovine rotavirus/isolated from CBNU-2 and 75 times passaged on MA104 cells), respectively, by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequenced and compared. The sequence data indicated that the NSP4 genes of bovine rotavirus (BRV) were 751 bases in length and encoded one open reading frame of 175 amino acids beginning at base 42 and terminating at base 569. Differences in nucleotide sequence between CBNU-2 and CBNU-1 were observed at 6 positions (base 274, 296, 391, 394, 396 and 579). NSP4 gene of BRV exhibited a high degree of nucleotide (90% and 94%) and amino acid sequence (91% and 97%) homology with those of SA11 and UK but a low degree of nucleotide (77% and 79%) and amino acids sequence (81% and 85%) homology with those of Wa and OSU.