• Journal of fish pathology
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• v.8 no.1
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• pp.31-36
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• 1995

Hc nuclear polyhedrosis virus DNA genome was digested with EcoRI endonuclease, these partial fragments were recombined into the pUC8 plasmid vector and transformed in E. coli JM 83 cell. The genome DNA has 24 EcoRI fragments and 12 fragments of them were subcloned. The four recombinants were named as eNP-O, eNP-Q, eNP-R and eNP-S. The expression of foregin gene of the recombinants was investigated by analysing protein patterns on the SDS-PAGE. The eNP-O, eNP-Q and eNP-R were expressed a different weight of protein as comparision with potypeptide bands of E. coli JM 83 host cell.

• Preventive Nutrition and Food Science
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• v.9 no.4
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• pp.324-329
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• 2004

In order to investigate the molecular mechanism of $\gamma$-aminobutyric acid (GABA) production in lactic acid bacteria, we cloned a glutamate decarboxylase (GAD) gene from Lactobacillus plantarum using polymerase chain reaction (PCR). One PCR product DNA was obtained and inserted into a TA cloning vector with a T7 promoter. The recombinant plasmid was used to transform E. coli. The insertion of the product was con­firmed by EcoRI digestion of the plasmid purified from the transformed E. coli. Nucleotide sequence analysis showed that the insert is a full-length Lactobacillus plantarum GAD and that the sequence is $100\%$ and $72\%$ identical to the regions of Lactobacillus plantarum GAD and Lactococcus lactis GAD sequences deposited in GenBank, accession nos: NP786643 and NP267446, respectively. The amino acid sequence deduced from the cloned Lactobacillus plantarum GAD gene showed $100\%$ and $68\%$ identities to the GAD sequences deduced from the genes of the NP786643 and NP267446, respectively. To express the GAD protein in E. coli, an expression vector with the GAD gene (pkk/GAD) was constructed and used to transform the UT481 E. coli strain and the expression was confirmed by analyzing the enzyme activity. The Lactobacillus plantarum GAD gene obtained may facilitate the study of the molecular mechanisms regulating GABA metabolism in lactic acid bacteria.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.153-156
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• 2000

Xylan, the major hemicellulose component of many plants, occurs naturally in a partially acetylated form and lignin, the most resistant component in plant cell wall degradation, is also attached to ${\beta}-1,4-linked-D-xylose$ backbone through the ester linkage. Esterases are required to release the esterified substituent and acetyl esterases are important in the complete degradation of acetylated polysaccharides, like pectins and xylans. The gene(Axe) encoding acetyl xylan estarase(AXE) was isolated from genomic ${\lambda}$ library from Aspergillus ficuum. Nucleotide sequencing of the Axe gene indicated that the gene was separated with two intervening sequences and the amino acid sequence comparison revealed that it was closely related to that from A. awamori with the 92 % indentity. Heterologous expression of AXE was conducted by using YEp352 and Saccharomyces cerevisae 2805 as a vector and host expression system, respectively. The Axe gene was placed between GAL1 promoter and GAL7 terminator and then this recombinant vector was used to transform S. cerevisiae 2805 strain. Culture filtrate of the transformed yeast was assayed for the presence of AXE activity by spectrophotometry and, comparing with the host strain, four to five times of enzyme activity was detected in culture filtrate of transformed yeast.

• Proceedings of the Korean Society of Life Science Conference
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• 2008.10a
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• pp.55.1-55.1
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• 2008

• Proceedings of the Korean Society of Life Science Conference
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• 2007.10a
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• pp.109.1-109.1
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• 2007

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• Journal of Microbiology and Biotechnology
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• v.21 no.12
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• pp.1294-1298
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• 2011

A metK gene encoding S-adenosyl-L-methionine synthetase was cloned from the non-Streptomyces actinomycetes, Actinoplanes teichomyceticus ATCC31121. In order to evaluate the effect of the metK expression on antibiotic production in actinomycetes, an expression vector harboring the metK gene was constructed and introduced into Streptomyces lividans TK24 and A. teichomyceticus, and the antibiotic production of the exconjugants was assessed. As a result, it was determined that the expression of metK induced 17-fold and 2.2-fold increases in actinorhodin production from S. lividans TK24 and teicoplanin production from A. teichomyceticus, respectively, compared with the control strains.

• Korean Journal of Microbiology
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• v.25 no.3
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• pp.165-172
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• 1987

An expression vector in a mammalian cell was constructed using the origin of replication (OR) and the promoters of SV40. The plasmid pSVOE was constructed by inserting SV40 DNA fragment (1, 118bp) containing SV40 OR and promoters into pBR322-1, and then a multiple cloning sequence was inserted at the immediate downstream of the late promoter of SV40 in the pSVOE vector. The plasmid was named pSVML. As a selection marker, thymidine kinase gene of herpes simplex virus with its promoter was inserted into EcoRI site of pSVML and the recombinant was named pSVML-TKp. To test the expression capacity of foreigen gene inserted at the multiple cloning site of pSVML, the thymidine kinase gene without its own promoter was inserted at the BamHI site of pSVML. The recombinant was named pSVML-TK. These plasmids, pSVML-TKp and pSVML-TK, were transfected into COS cells with calcium phosphate precipitation method. The thymidine kinase activity was significantly increased in both transfected cells.

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.90-90
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• 2002

In this study, we report the cloning of the porcine UPII genomic DNA, which contains a putative full-length open reading frame encoding the UPII protein. A comparison of the porcine UPII gene coding sequence with the previously published mouse UPII sequence demonstrates that only the exon sequences are partially conserved. Northern and immunohistochemical analyses show that the porcine UPII gene is expressed only in the urothelium and that the protein specifically localizes to urothelial superficial cells. (omitted)

• Proceedings of the Korean Society of Veterinary Clinics Conference
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• 2008.10a
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• pp.162-162
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• 2008

• BMB Reports
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• v.32 no.5
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• pp.497-501
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• 1999

Biotinylation of recombinant proteins is a powerful tool for the detection and analysis of proteins of interest in a large variety of assay systems. The recent development of in vivo biotinylation techniques in E. coli has opened new possibilities for the production of site-specifically biotinylated proteins without the need for further manipulation after the isolation of the recombinantly expressed proteins. In the present study, a novel vector set was generated which allows the convenient cloning and expression of proteins of interest fused with an N-terminal in vivo biotinylated thioredoxin (TRX) protein. These vectors were derived from the previously reported pBIOTRX vector into which was incorporated part of the pBluescript II+phagemid multiple cloning site (MCS), amplified by PCR using a pair of sophisticated oligonucleotide primers. The functionality of these novel vectors was examined in this system by recombinant expression of rat transforming growth factor-$\beta$. Western-blot analysis using TRX-specific antibodies or peroxidase-conjugated streptavidin confirmed the successful induction of the fusion protein and the in vivo conjugation of biotin molecules, respectively. The convenience of molecular subcloning provided by the MCS and the effective in vivo biotinylation of proteins of interest makes this novel vector set an interesting alternative for the production of biotinylated proteins.