• 한국미생물·생명공학회지
• /
• 제22권5호
• /
• pp.485-490
• /
• 1994

A restriction endonuclease, PsyI, has been isolated from Pseudomonas syringae pv. pha- seolicola, and its catalytic properties have been studied. This enzyme was purified through strepto- mycin sulfate and ammonium sulfate fractionation, phosphocellulose Pll, DEAE-cellulose, hydroxy- apatite and Sephadex G-100 column chromatography. It's molecular weight was about 50,000 dalton as determined by 7.5% polyacrylamide gel electrophoresis containing 0.1% SDS. In catalytic proper- ties, PsyI shows stable at wide ranges of pH between 7.0 and 10.0, of temperature between 30$\circ$C and 37$\circ$C, and its thermal stability is between 25$\circ$C, and 45$\circ$C, at the presence Of 10 mM MgCl$_{2}$-PsyI essentially require Na salt for enzyme reaction, is rather inhibited in the high Na salt concent- ration. The presence of 2-mercaptoethanol is absolutely required for the enzyme activity. This endonuclease, PsyI was determined to be an isoschizomer of SalI from the results of the restriction mapping and DNA sequencing.

• Asian-Australasian Journal of Animal Sciences
• /
• 제24권9호
• /
• pp.1274-1281
• /
• 2011

Dietary protein restriction affects lipid metabolism in rats. This study was performed to determine the effect of a low protein diet on hepatic lipid metabolism and insulin sensitivity in growing male rats. Growing rats were fed either a control 20% protein diet or an 8% low protein diet. Feeding a low protein diet for four weeks from 8 weeks of age induced a fatty liver. Expression of acetyl-CoA carboxylase, a key lipogenic enzyme, was increased in rats fed a low protein diet. Feeding a low protein diet decreased very low density lipoprotein (VLDL) secretion without statistical significance. Feeding a low protein diet down-regulated protein expression of microsomal triglyceride transfer protein, an important enzyme of VLDL secretion. Feeding a low protein diet increased serum adiponectin levels. We performed glucose tolerance test (GTT) and insulin tolerance test (ITT). Both GTT and ITT were increased in protein-restricted growing rats. Our results demonstrate that dietary protein restriction increases insulin sensitivity and that this could be due to low-protein diet-mediated metabolic adaptation. In addition, increased adiponectin levels may influences insulin sensitivity. In conclusion, dietary protein restriction induces a fatty liver. Both increased lipogenesis and decreased VLDL secretion has contributed to this metabolic changes. In addition, insulin resistance was not associated with fatty liver induced by protein restriction.

• 미생물학회지
• /
• 제23권2호
• /
• pp.122-128
• /
• 1985

Bacillus thuringiensis serovar israelensis 4Q1 contains 8 different covalently closed-circular (CCC) plasmids of molecular weight 204, 267, 109, 103, 16, 7.6, 6.4, and 5.0kb. The three smallest plasmids, designated pBti6, and pBti8 may prove to be useful as cloning vectors because of thier size and ease of isolation. The three plasmids were incubated separately with 9 different restriction enzymes and 7 of the enzymes tested cleaved one or more of the plasmids. Plasmid pBti6 has a single site for Bg1 II, Pst I and Pvu II, two sites for Bc1 I and Eco RI, and five sites for Hind III. Plasmid pBti7 has a single site for Bam HI and Pst I, two sites for Hind III, and three sites for PvuII. Plasmic pBti8 has a single site for Bam HI, BelI and Hind III, two sites for Eco RI, and three sites for Bgl II and Pvu II. Composite restriction enzyme maps for pBti6, pBti7 and pBti8 were constructed. The sites of restriction enzyme cleavage were determined by single, double and partial digests of the plasmid DNA. All the restriction sites were aligned relative to the single Bgl II(pBti6), Pst I(pBti7) or Hind III(pBti8) site, respectively.

• Journal of Microbiology
• /
• 제43권5호
• /
• pp.417-423
• /
• 2005

In this study, the nematode-trapping fungus, Monacrosporium sphaeroides, was transformed with a plasmid harboring the hygromycin B phosphotransferase gene, via restriction enzyme-mediated integration (REMI). Frequencies of up to 94 transformants ${\mu}g^{-1}$ per linearized plasmid DNA were obtained by optimizing the PEG concentration, as well as the category and quantity of the added restriction enzyme. $90\%$ of the transformants were determined to be stable for drug resistance when 20 randomly selected transformants were tested. Southern analyses revealed that the transforming DNA was integrated into the M. sphaeroides genome either with or without rearrangement. Five mitotic stable mutant strains were obtained using this approach, all of which had been altered with regard to sporulation capacity and pathogenicity toward nematodes. Southern blot analyses of the five mutants revealed that foreign plasmid DNA had integrated into the genome. Three of the mutants, Tms2316, Tms3583 and Tms1536, exhibited integration at a single location, whereas the remaining two, Tms32 and Tms1913, manifested integration at double or multiple locations. Our results suggest that the transformation of M. sphaeroides via REMI will facilitate insertional mutagenesis, the functional analysis of a variety of genes, and the tagging or cloning of genes of interest.

• 한국작물학회지
• /
• 제44권3호
• /
• pp.253-255
• /
• 1999

Reverse transcription and polymerase chain reaction (RT-PCR) assay was used to detect SMV strains. A pair of oligonucleotide primers were designed to include the cylindrical inclusion (CI) coding region between 4,176 to 5,560 nt. Amplification from the total RNA extracted from infected plants with SMV yielded a 1,385 bp DNA fragment. RT-PCR was shown to be $10^3$ times more sensitive than the ELISA assay and it could detect a virus in $10^{-6}$ dilution. Restriction enzyme analysis of RT- PCR products using EcoR I showed that SMV isolates were classified into six groups according to the patterns of restriction fragments.

• 약학회지
• /
• 제36권3호
• /
• pp.255-258
• /
• 1992

The clinical isolate Staphylococcus aureus SA8 was resistant to tetracycline(Tc) and harboured a plasmid pKH1(24.82 kb). pKH1 was shown by curing and by transformation to specify resistance to Tc. The cleavage map of a pKH1 was determined by restricction enzyme mapping techniques. Cleavage map is given for BglII, EcoRI, HpaII, PvuII and SalI. Restriction endonuclease BamHI, BglI, BstEII, HpaI, PstI, and XhoI have no sites on this plasmid. HaeIII, XbaI, and HindIII have 5, 6, 14 sites, respectively.

• Journal of Microbiology
• /
• 제34권4호
• /
• pp.334-340
• /
• 1996

The enzyme, cis,cis-muconate lactonizing enzyme has been proposed to play a key role in the $\beta$-ketoadipate pathway of benzoate degradation. A 3.2-kb EcoRI fragment termed as pRSU2, isolated from a Pseudomonas cepacia genomic library was able to complement the catB defective mutant. Several relevant restriction enzyme sites were determined within the cloned fragment. In Pseudomonas putida SUC2 carrying pRSU2, the enzyme activity was relatively higher than those of the induced or partially induced state of wild type P. putida PRS2000. It was probably due to higher expression of P. cepacia catB in P. putida PRS2000. It was probably due to higher expression of P. cepacia catB in P. putida. One possible interpretation of these results is that the catB promoter in P. cepacia is recognized within P. putida, resulting in the almost same expression level.

• 한국식품영양과학회지
• /
• 제28권2호
• /
• pp.471-476
• /
• 1999

The effects of total dietary restriction(100% restriction of energy intake) on thiobarbituric acid reactive substance(TBARS) contents and intracellular antioxidant enzymes activities in the liver and kidney of young male Sprague Dawley rats were studied. The TBARS contents were reduced in both liver and kidney, up to 77% and 79% of the control rats, fed ad libitum, respectively at 7 days after dietary restriction . Superoxide dismutase(SOD) activities in the liver and kidney of rats were increased significantly by total dietary restriction. However, the activity of catalase in kidney was decreased 27% at 6 days after dietary restriction, but this enzyme activity did not change in liver. The changes of glutathione peroxidae(GSHPx) and catalase activities in the liver and kidney of rats with dietary restriction were not significant. These result suggested that dietary restriction reduce the free radical induced by tissue damage, as determined by TBARS content, in both the liver and kidney but the changes of activities of antioxidant enzymes may not be a contributory factor in reducing oxidative damage to tissue.

• 한국자원식물학회지
• /
• 제21권1호
• /
• pp.41-46
• /
• 2008

차나무의 분자학적 유연관계 및 유전적 다양성을 알아보기 위하여 한국에서 자생하는 차나무 집단 21개 지역 차나무에 대하여 DFR 유전자 부위를 이용하여 PCR-RFLP분석을 하였다. DFR 4+5 primer 쌍을 이용하여 증폭결과 DFR유전자의 크기는 약 1.4kb에서 PCR산물을 획득하였다. PCR산물에 제한효소 Hpa II 및 Mse I를 이용하여 RFLP분석 결과 차나무 집단간 또는 집단내 차나무간의 유전적 다양성을 보였다. Hpa II 제한효소를 이용한 RFLP 밴드패턴은 3가지 형태로 구분되었으며, 같은 차나무 집단내에서도 유전적 다양성이 나타났다. Mse I 제한효소를 이용한 밴드패턴은 6가지의 형태로 다양성을 보였으며, 웅포집단 차나무의 경우 집단 내 다양성이 2가지 형태로 나타나 같은 집단내에서의 유전적 변이는 적은 것으로 보인다. 본 연구에서 사용한 2가지의 제한효소의 결과 차나무의 집단간 또는 같은 집단내의 차나무간에서 유전적 다양성을 확인할 수 있었다.

• 한국가축번식학회지
• /
• 제27권2호
• /
• pp.97-102
• /
• 2003

본 연구는 분만시 동복 신생자돈의 제대혈에서 추출한 genomic DNA를 PCR-RFLP 기법을 이용하여 농가수입증대를 위하여 육질이 불량한 PSS 돼지를 판별하는 방법을 개발하기 위한 기초실험으로써 그 결과를 요약하면 다음과 같다. 자돈 제대에서 혈액 genomic DNA를 추출하여 PCR에 의하여 증폭된 ryanodine receptor gene 영역의 산물은 자돈의 제대혈에서 1.8kb의 길이로 증폭되었음을 확인하였다. 제대혈에서 추출된 DNA 의 PCR 증폭 단편을 가지고 Hha I 제한효소로 digest 하여준 결과에서 PSS 돼지는 Yorkshire 종에서 출현하지 않았으나, Landrace 종과 Crossbred 종에서 각각 4.76%와 7.14%로 교잡종(LYD 또는 YLD)에서 더욱 많이 출현되었다. 이상의 결과로 볼 때 분만시 신생자돈의 제대혈을 채취하여 PSS 돼지를 조기에 선발하면 스트레스 감소는 물론 혈액채취의 간편성을 기대할 수 있을 것으로 판단된다.