• 미생물학회지
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• 제30권3호
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• pp.232-238
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• 1992

Campylobacter jejuni 에 열처리를 했을 때 그들의 생존성 및 열충격 단백질 합성의 양상과 더불어, dnaK 와 groESL 유전자를 이용하여 C. jejuni 의 열충격 유전자를 검출하여 그 특성을 E. coli 의 열충격 유전자와 비교하였다. C. jejuni 의 열충격 단백질은 48.deg.C 에서 가장 잘 발형되었으며, 48.deg.C 에서 30 분간의 처리중 세포들의 생존율은 떨어지지 않았다. C. jejuni 의 열충격 단백질로서의 Hsp90, Hsp66, Hsp60 이 합성되는 것을 SDS-PAGE 및 방사선사진법을 통해 확인하였다. dnaK 와 groESL 을 DNA 탐침자로 이용하여 Southern hybridization 한 결과, C. jejuni 의 열충격 유전자도 groESL 과 dnaK 유전자와 상동성을 가진 염기서열을 가지고 있었으나, 두 균주사이에는 열충격유전자를 내포하고 있는 DNA 상에서 제한효소의 절단부위에 차이가 있었다.

• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.663-669
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• 2000

The fur (ferric uptake regulation) expression vector pMON2064 was modified to produce a Fur-fusion expression vector. A kinker site, factor Xa cleavage site, and several restriction endonuclease sites were introduced to facilitate easy cloning and isolating of the fusion protein. The resulting fusion expression vector, pMONSTER, was then used to make fusion expression vector, pMONSTER, was then used to make fusion proteins with $\beta$-galactosidase and the protease of the human immunodeficiency virus type 1 (HIV-1 PR). Strain SW4020 harboring the Fur $\beta$-galactosidase fusion vector produced blue colonies on a 5-bromo-4-chloro-3-indolyl-$\beta$-D-galactoside plate and the resulting 133 kDa fusion protein reacted with an anti-Fur antibody. The strain harboring the Fur-HIV-1 PR fusion vector produced a 29 kDa fusion protein, which also reacted with an anti-Fur antibody. The Fur-HIV-1 PR fusion protein was purified by a single column application that was designed to isolate the Fur protein. The purified Fur-HIV-1 PR fusion protein digested with factor Xa cleaved a recombinant Gag protein to release smaller fragments, including a p24 capsid protein. The Fur-HIV-1 PR fusion protein itself did not exhibit any proteolytic activity.

• Journal of Microbiology and Biotechnology
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• 제19권12호
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• pp.1620-1627
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• 2009

The design and expression of an antihypertensive peptide multimer (AHPM), a common precursor of 11 kinds of antihypertensive peptides (AHPs) tandemly linked up according to the restriction sites of gastrointestinal proteases, were explored. The DNA fragment encoding the AHPM was chemically synthesized and cloned into expression vector pGEX-3X. After an optimum induction with IPTG, the recombinant AHPM fused with glutathione S-transferase (GST-AHPM) was expressed mostly as inclusion body in Escherichia coli BL21 and reached the maximal production, 35% of total intracellular protein. The inclusion body was washed, dissolved, and purified by cation-exchange chromatography under denaturing conditions, followed by refolding together with size-exclusion chromatography and gradual dialysis. The resulting yield of the soluble GSTAHPM (34 kDa) with a purity of 95% reached 399 mg/l culture. The release of high active fragments from the AHPM was confirmed by the simulated gastrointestinal digestion. The results suggest that the design strategy and production method of the AHPM will be useful to obtain a large quantity of recombinant AHPs at a low cost.

• Asian-Australasian Journal of Animal Sciences
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• 제20권6호
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• pp.827-831
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• 2007

Myostatin is a member of the transforming growth factor-${\beta}$(TGF-${\beta}$ super-family. It acts as a negative regulator for skeletal muscle growth. Myostatin mutations are characterized by a visible, generalized increase in muscle mass in double muscled cattle breeds. To understand the biochemistry and physiology of the Chinese Yellow bovine myostatin gene, we report here for the first time expression of the gene in Escherichia coli (E. coli). Primers of the myostatin gene of Chinese Yellow Cattle were designed on the basis of the reported bovine myostatin mRNA sequence (Gen-Bank Accession No. NM005259) and optimized for E. coli codon usage. XhoI and EcoRI restriction enzyme sites were incorporated in the primers, and then cloning vector and expression vector were constructed in a different host bacterium. The expressed protein had a molecule mass of about 16 kDa as determined by SDS-PAGE under reducing conditions. The expressed protein reacted specifically with myostatin monoclonal antibody on immunoblots. Our studies should lead to the investigation of the differences in myostatin genes of various cattle and could benefit human health and food animal agriculture.

• Journal of Microbiology and Biotechnology
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• 제9권1호
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• pp.91-97
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• 1999

In this work, a 1.9-kb region of enterococcal plasmid p703/5 was isolated and the nucleotide sequence analysis of the region was performed. One major open reading frame (ORF) was identified encoding a polypeptide of 28 kDa. Database comparisons suggested that the protein showed some homology with other bacterial RepA proteins. Upstream of the ORF, a potential dnaA box, AT-rich region and 22-bp tandemly repeated sequences (DNA iterons), a feature typical for many replication ori sites, were recognized. Deletion analysis using Exonuclease III and several restriction enzymes indicated that the three elements and the gene product from the ORF were essential for replication and that the minimum unit of DNA required for replication resided on the 1.2-kb AvaII subfragment. Thus, this gene product was referred to as RepA.

• 한국미생물·생명공학회지
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• 제22권1호
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• pp.37-44
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• 1994

The homology among the genes coding for degradation of bipheny(BP) and 4-chlorobiphenyl(4CB) was comparatively analyzed by Southern hybridization in several BP/4CB degrading bacterial strains. As the hybridization results of their genomic DNAs with pcbABCD as the DNA probe, the group of Pseudomonas sp. DJ-12. P08 and P27 strain was separated by the group of P20 and P1242 strains. The P. pseudoalcaligenes KF707 showed the hybidization signal which was homologous to the group of DJ-12, but they had different restriction endonuclease sites. The pcbAB genes in pCUl recombinant plasmid from Pseudomonas sp. DJ-12 appeared to be homologous to pchAB genes in pKTF20 cloned from P. pseudoalcaligenes KF707, but the C genes in both strains were not homologous. The bphABC in pKTF20 showed the signals homologous to the cbp ACB in pAW6194 cloned from P. putida OU83, but homologous signal was not found botween the pcbABCD genes in pCUl and the cbpADCB genes in pAW6194 recombbinant plasmid.

• 한국미생물·생명공학회지
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• 제22권1호
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• pp.31-36
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• 1994

From the pSKH201 plasmid which had been previously cloned in our laboratory, a 3.0kbp insert DNA encoding the alkaline phosphatase of Kluyveromyces fragilis was cleaved with several restriction endonucleases and ligated int the appropriate sites of M13mp18/19 vectors and sequenced by Sanger's dideoxy chain termination method. The sequence contained a 1,638 bp open reading frame(ORP) whose similarities in nucleotide, when compared with those of Saccharomyces cerevisiae and Escherichia coli by GENETYX program, were found to be 61% and 46%, respectively. The deduced amino acid sequence consists of 546 amino acids and contains several homologous regions in the alkaline phosphatases of E. coli, S.cerevisiae and human placenta.

• 한국균학회지
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• 제17권1호
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• pp.35-38
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• 1989

팽나무버섯 균사체 DNA를 대장균 프라스미드인 PBR322를 이용하여 gene library를 작성 하였다. 팽나무버섯에서 ${\beta}-isopropyl$ malate dehydrogenase 유전자 클론을 얻었으며 이 클론은 대장균 Leucine 요구성 균주를 complementation 시켰다. 이 클론의 팽나무버섯 DNA 크기는 약 lKb였으며 BamH1과 Ava 1 제한효소 절단부위를 지니고 있었다.

• 한국전자거래학회지
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• 제6권2호
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• pp.87-99
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• 2001

Along with the rapid emergence of the Internet and e-commerce, online auctions are hitting the spotlights. The inconveniences found in off-line auctions, such as time and place restriction and limited number of items, are solved in the online auction. However, not so many auction sites have integrated auction information systems, which monitor the present status of auctions, resulting in greater inconvenience for the online auction users. Moreover, there is no auction site that suggests the appropriate starting or closing price that is useful for users when they make, their bids, What the online auction users need is an auction system that can solve such problems. This study is purported for solving the problems by designing and implementing a real time auction system that applies the comparison search functions and the agent functions. In other words, an integrated database system using a bidder-oriented agent for providing information is built so that the users can search and compare the information on the item they are interested in and make a faster and more accurate purchase. Also the appropriate starting and closing prices are offered to the sellers and bidders through the integrated system for a closer and more accurate comparison and analysis of the prices. For future work, the product recommendation service, which accurately reflects the bidding patterns, and the methods for studying the multi bidding pattern will be applied to the suggested system to realize a real time auction information system that supports CRM(Customer Relationship Management) .

• 대한의생명과학회지
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• 제8권1호
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• pp.29-37
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• 2002

Listeria monocytogenes poses an increasing health risk, which in part is due to increasing health risk, consumption of ready-to-eat food products and the introduction of increasing numbers of food products from regions with different dietary habits. L. monocytogenes can be present in meat, shellfish, vegetables, unpasteurised milk and soft cheese and poses a risk if food containing these products is stored at refrigeration temperature and is not properly heated before consumption, as L. monocytogenes is psychrophilic. Amplified-fragment length polymorphism (AFLP) analysis is the method of genotypic techinique in which adaptor oligonucleotides are ligated to restriction enzyme fragments and then used as target sites for primers in a PCR amplification. The amplified fragments are electrophoretically separated to give strain-specific band profiles. Single-enzyme approach that did not require costly equipment or reagents for the fingerprinting of strains of Listeria monocytogenes was developed. Single-enzyme amplified fragment length polymorphism (SE-AFLP) analysis was used to perform species and strain identification of Salmonella, Shigella, Yersinia and E. coli. By careful selection of AFLP primers, it was possible to obtain reproducible and sensitive identification to strain level. The AFLP patterns of L. monocytogenes are divided by the kinds of specimens in which were isolated. SE-AFLP fragments can be analyzed using standard gel electrophoresis, and can be easily scored by visual inspection, due to the low complexity of the fingerprint obtained by this method. These features make SE-AFLP suitable for use in either field or laboratory applications.