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PCR을 이용한 식품 내 Salmonella 균주의 신속 검출방법
정상훈,김묘영,김현중,김태운,유상렬,김해영,Jung, Sang-Hun,Kim, Myo-Young,Kim, Hyun-Joong,Kim, Tae-Woon,Ryu, Sang-Ryeol,Kim, Hae-Yeong 한국응용생명화학회 2003 한국농화학회지 Vol.46 No.3
여러 종류의 식품에서 Salmonella 균주를 손쉽고 빠르게 검출하기 위하여, Salmonella 장독소 유전자(stn)를 기초로 제작한 primer (STN1, STN2)를 이용하여 PCR을 수행한 결과 617 hp의 특이적인 DNA단편을 얻을 수 있었다. PCR 민감도는 순수 배양한 균체에서 추출한 template DNA는 1 pg까지 검출이 가능하였고, 직접 균체를 template로 이용한 경우에는 $10^2\;cells$까지 검출이 가능하였다. Salmonella typhimurium을 인위적으로 접종시킨 식품에서는 식품 1 g당 $10^3{\sim}10^4$ cells까지 검출할 수 있었다. 이러한 결과는 PCR을 이용하여 Salmonella에 오염된 식품에서 이들 균주를 간편하고 신속하게 검출할 수 있을 것이다. This study was carried out to investigate the simple and rapid detection of Salmonella species in different kinds of food using PCR method. The specific primer sets (SIN1 and SIN2) was designed and utilized to amplify a 617 bp DNA fragment from salmonella species. The sensitivity of PCR was 1 pg of purified template DNA or $10^2$ cells from pure culture. The detection limit of Salmonella typhimurium on agarose gel electrophoresis was $10^3{\sim}10^4$ cells/g in the artificially contaminated food samples. These results suggested that this simple method could be applied to industrial fields for detection of Salmonella species in food.
한국에서 분리된 병원성 Salmonella 균주의 장독소 유전자(stn) 분포와 발현조절 기작
유상렬 서울대학교 농업개발연구소 2000 농업생명과학연구 Vol.4 No.-
Role of enterotoxin from Salmonella in pathogenesis is not known. Enterotoxin gene from Salmonella typhimurium (stn) encodes a 29 kDa toxin that has no homology to any other known enterotoxins. Expression of stn is enhanced upon contact with epithelial cell but not all strains having the stn gene express Stn. Based on PCR analysis, we found that all 36 clinical strains of Salmonella isolated in Korea tested carried the stn gene. To understand the regulation of the stn transcription, the expression of stn was studied in vitro. The expression of stn was inhibited by cAMPㆍCRP complex by about 50%.
Mlc 조절단백질이 대장균의 pts 유전자 발현에 미치는 영향
유상렬 서울대학교 농업개발연구소 1999 농업생명과학연구 Vol.3 No.-
Products of the pts operon of Escherichia coli have multiple physiological roles and the operon is controlled by two promoters, P0 and P1. Expression of the pts P0 promoter that is increased during growth in the presence of glucose is also activated by CRP-cAMP. The effects of the Mlc on the pts P0 expression were studied. In vivo transcription assay using wild type and Mlc strains grown in the presence and absence of glucose indicate that Mlc negatively regulates expression of P0, and Mlc-dependent repression is relieved by glucose in the growth medium. In vitro transcription assay using purified Mlc showed that Mlc repressed transcription from the P0 specifically.
Analysis of Salmonella pathogenicity island (SPI) 2 gene expression
Ryu, Sang-Ryeol 서울대학교 농업개발연구소 2001 농업생명과학연구 Vol.5 No.-
Salmonella pathogenicity island 2 (SPI2) encodes a putative type Ⅲ secretion system necessary for replication inside macrophages systemic infection in animals. The genes for the secretion system such as ssaH are transcribed preferentially after Salmonella enters host cells. The transcriptional organization of 3 genes of SPI2 was investigated, which encode components of one of two type Ⅲ secretion apparatus of Salmonella typhimurium. ssaH, I, J constitute one operon of 1.5 kb and the promoter of this operon lies upstream of ssaH. The transcriptional regulation of ssaH (operon) was studied by primer extension analysis. It was found that stationary growth phase-specific transcription of ssaH is most efficient in the presence of OmpR (pleiotropic regulator of Salmonella virulence genes), InvF, SsrB (local regulator of SPI1 and 2), IHF, and Fis (nucleoid associated proteins). Furthermore, we constructed two kinds of reporter plasmid to know whether the ssaH expression is specific to Salmonella. In this study, it was found that SsrB was prerequisite of ssaH expression because a ssaH was not expressed in E. coli which is absent of SPI.