• 한국독성학회:학술대회논문집
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• 한국독성학회 2003년도 추계학술대회
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• pp.86-86
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• 2003

Gene therapy is a medical intervention based on modification of the genetic material of living cells. Gene transfer usually conducted using bacterial plasmid DNA and/or virus vector to express a specific protein. Gene transfer medicinal products classified as naked nucleic acid, complexed nucleic acid or non-viral vectors, viral vector, and genetically modified cells according to biological origin.(omitted)

• 한국어류학회지
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• 제19권2호
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• pp.83-87
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• 2007

어류 에드워드감염증에 대한 예방 재조합 ghost 백신을 개발하기 위한 연구의 일환으로 어류 병원성 세균인 Edwardsiella trada를 대상으로 플라스미드 형질전환을 실시하고 형질전환 안정성을 평가하였으며, 형질 도입된 재조합 ghost 세균의 E-lysis 유전자 발현을 분석하였다. E. tarda를 대상으로 한 ghost 유도는 대장균에 비해 상대적으로 장시간의 반응 시간이 요구되며 lysis의 개시가 지연된 점을 고려 시 발현된 E protein의 용해 능력 또는 E-gene의 전사발현 양이 E. tarda에서 다소 약화되는 것으로 나타났다. 그러나 대장균에 비해 ghost 유도 속도가 다소 낮음에도 불구하고 반응이 완성되었을 시점에서의 E. tarda의 ghost 효율은 대장균과 전혀 차이가 없이 99.99% 이상의 유도효율을 나타내었다.

• Archives of Pharmacal Research
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• 제21권3호
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• pp.305-309
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• 1998

Human cytochrome P450 1A2 is one of the major cytochrome P450s in human liver. It is known to be capable of activating a number of carcinogens such as arylamines and heterocyclic amines. In order to develop the new bacterial mutagenicity test system with human P450, a full length of human P450 1A2 cDNA inserted into pCW bacterial expression vector was introduced to Escherichia coli WP2 uvrA strain which is a well-known E. coli strain for bacterial reverse mutagenicity assay. Expressed human P450 1A2 showed typical P450 hemoprotein spectra. Maximum expression was achieved at 48 hrs after incubating at $30^{\circ}C$ in terrific broth containing ampicillin, IPTG and other supplements. High level expression of P450 1A2 in E. coli WP2 uvrA membranes was determined in SDS-PAGE. The well-known mutagens 2-aminoanthracene and MElQ increased the revertant colonies of E. coli WP2 uvrA expressing human P450 1A2 without an exogenous rat hepatic post-mitochondrial supernatant (S9 fraction) in a dose-dependent manner. The results show that the functional expression of human P450 in bacterial mutagenicity tester strain will provide a useful tool for studying the mechanism of the mutagenesis and carcinogenesis of new drugs and environmental chemicals.

• 한국지능시스템학회:학술대회논문집
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• 한국퍼지및지능시스템학회 2006년도 춘계학술대회 학술발표 논문집 제16권 제1호
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• pp.22-34
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• 2006

o GA-BF approach for improvement of learning and optimization in GA o GA-BF has better response on various test functions o Satisfactory PID controller tuning in AVR, motor vector control systems o Potentially useful in many practically important engineering optimization problems

• Journal of Microbiology and Biotechnology
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• 제12권4호
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• pp.706-709
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• 2002

Bacterial bioluminescence has been known to be a highly valuable reporter system for its potential application as an effective and simple environmental monitoring method for toxic compounds. In this short report, we constructed a streptomycetes-Escherichia coli shuttle vector-containing bioluminescence system and evaluated its potential application for toxic compounds monitoring. The luxAB biolurninescence genes from Vibrio harveyi were cloned into a streptornycetes-E. coli shuttle vector (named pESK004) and functionally expressed in Streptomyces lividans. The recombinant S. lividans containing pESK004 exhibited an optimal biolurninescence at the optical density ($OD_{600\;nm}$) of 0.4-0.5 and aldehyde concentration of 0.005%. When the recombinant bioluminescence streptomycetes was exposed to a toxic compound such as heavy metals, chlorinated phenols, or pesticides, the bioluminescence was decreased proportionally to the concentration of toxic compound in the assay mixture. The $EC_{50}$ (effective concentration to decrease 50% of the bioluminescence prior to exposure) values in the recombinant biolurninescence streptomycetes for mercury, 2,4-dichlorophenol, and malathion were measured at 2.2 ppm, 144.0 ppm, and 82.4 ppm, respectively. The degree of sensitivity and specificity pattern toward these toxic compounds characterized in this recombinant bioluminescence streptomycetes were unique when compared with previously reported bacterial bioluminescence systems, and this revealed that a recombinant bioluminescence streptomycetes might provide an alternative or complementary system for potential environmental monitoring.

• Journal of Microbiology and Biotechnology
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• 제13권6호
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• pp.872-880
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• 2003

Three vectors, pSG1, 2, and 3, which facilitate in vivo expression technology (IVET) in Gram-negative bacteria, were developed. Vectors pSG1and 2 are derivatives of ColE1, and pSG3 is a derivative of an R6K replicon. These vectors contain oriT sites that allow mobilization when the RK2 Tra functions are provided in trans. These vectors contain promoterless lacZ (pl-lacZ) and promoterless aph (pl-aph) transcriptionally fused together, which allow qualitative and quantitative measurements of the expression of genes in the genome of bacterial cells. pSG1 and 3 contain gentamicin-resistance genes, and pSG2 carries a streptomycin-/spectinomycin-resistance gene, allowing for selection of recombinants generated by a single crossover between a library fragment cloned into a pSG vector and the identical region in the genome of a bacterial species from which the library fragment originated. These vectors were successfully applied to the generation of random fusions at high rates in the genomes of four representative Gram-negative bacteria. In addition, the expression level of ${\beta}-galactosidase$ and the degree of resistance to kanamycin in cells with fusions generated by these vectors were found to be linearly correlated, proving that these vectors can be used for IVET.

• 생명과학회지
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• 제22권2호
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• pp.177-183
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• 2012

Ralstonia solanacearum (Rs)에 의해 유발되는 풋마름병은 감자 재배 시 발병하는 주요 병 중의 하나이다. 감자에서 풋마름병 저항성관련 유전자를 찾기 위해 기존에 기능이 알려진 다른 가지과 작물의 기능 유사 유전체를 이용하여 StACRE (HM749652) 유전자를 분리하고 염기서열을 분석하였다. 분리한 StACRE의 발현양상을 분석하기 위해 병 저항성 유도 신호전달 물질인 SA와 풋마름병원균 Rs (KACC10722)를 처리한 감자에서 RNA를 추출하여 RT-PCR을 실시한 결과 이 유전자는 SA 처리에 의해 3시간 후부터, Rs에 의해서는 12시간 후부터 발현이 현저하게 증가하였다. 따라서, 감자에서 이 유전자의 생물학적인 기능을 분석하기 위해 Gateway System을 이용하여 과발현용 vector를 만든 후 과발현 형질전환 감자를 제작하고 풋마름병균인 Rs를 접종하여 병 저항성 기능을 검정 한 결과 대조구인 수미 감자에 비해 병 저항성이 증대하였다.

• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1180-1184
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• 2006

The capsule that surrounds Yersinia pestis cells is composed of a protein-polysacchride complex; the purified protein component is fraction I (F1) antigen. We report the cloning of the cafl gene and its expression in Escherichia coli using the vector pETl02/D-TOPO and the F1-specific monoclonal antibody. The recombinant F1 (rF1) antigen had a molecular size of 17.5 kDa, which was identical to that of the F1 antigen produced by Y. pestis. Recombinant F1 protein was found to react to polyclonal antiserum to Y. pestis Fl. Recombinant F1 was purified by ProBond purification system and induced a protective immune response in BALB/c mice challenged with up to 10$^5$ virulent Y. pestis. Purified rF1 protein was used in an ELISA to evaluate the ability of a method to detect antibodies to Y. pestis in animal sera. These results strongly indicated that the rF1 protein is a suitable species-specific immunodiagnostic antigen and vaccine candidate.

• 대한수의학회지
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• 제54권3호
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• pp.147-150
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• 2014

Q-fever is a vector-borne (Coxiella [C.] burnetii) zoonotic disease that is an increasing public health concern. To date, some research about Q-fever prevalence in dairy herds and human patients has been reported in Korea, but information about Korean native cattle is scarce. To measure the prevalence rates of C. burnetii in Korean native cattle, a total of 1,095 bovine serum samples collected during 2010~2013 were analyzed with an enzyme-linked immunosorbent assay. Sixty-eight heads of cattle were diagnosed as positive and while 19 heads were suspected (positive rate = 6.2%). Interestingly, Jeju province had a seropositivity rate six times greater than that of other provinces (18.9% vs. 3.2%). High seroprevalence might be caused by wide distribution of ticks in Jeju province compared to other regions. Based on these data, extensive monitoring of C. burnetii infection in cattle, tick distribution, and climate changes is required.

• 한국미생물·생명공학회지
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• 제29권1호
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• pp.1-11
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• 2001

Lactic acid bacteria (LAB) are widely used for various food fermentation. With the recent advances in modern biotechnology, a variety of bio-products with the high economic values have been produced using microorganisms. For molecular cloning and expression studies on the gene of interest, E. coli has been widely used mainly because vector systems are fully developed. Most plasmid vectors currently used for E, coli carry antibiotic-resistant markers. As it is generally believed that the antibiotic resistance markers are potentially transferred to other bacteria, application of the plasmid vectors carrying antibiotic resistance genes as selection markers should be avoided, especially for human consump-tion. By contrast, as LAB have some desirable traits such that the they are GRAS(generally recognized as safe), able to secrete gene products out of cell, and their low protease activities, they are regarded as an ideal organism for the genetic manipulation, including cloning and expression of homologous and heterologous genes. However, the vec-tor systems established for LAB are stil insufficient to over-produce gene products, stably, limiting the use of these organisms for industrial applications. For a past decade, the two popular plasmid vectors, pAM$\beta$1 of Streptococcus faecalis and pGK12 theB. subtilis-E. coli shuttle vector derived from pWV01 of Lactococcus lactis ssp. cremoris wg 2, were most widely used to construct efficient chimeric vectors to be stably maintained in many industrial strains of LAB. Currently, non-antibiotic markers such as nisin resistance($Nis^{r}$ ) are explored for selecting recombi-nant clone. In addition, a gene encoding S-layer protein, slp/A, on bacterial cell wall was successfully recombined with the proper LAB vectors LAB vectors for excretion of the heterologous gene product from LAB Many food-grade host vec-tor systems were successfully developed, which allowed stable integration of multiple plasmid copies in the vec-mosome of LAB. More recently, an integration vector system based on the site-specific integration apparatus of temperate lactococcal bacteriophage, containing the integrase gene(int) and phage attachment site(attP), was pub-lished. In conclusion, when various vector system, which are maintain stably and expressed strongly in LAB, are developed, lost of such food products as enzymes, pharmaceuticals, bioactive food ingredients for human consump-tion would be produced at a full scale in LAB.