• Journal of Food Hygiene and Safety
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• v.26 no.4
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• pp.296-301
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• 2011

This study was to evaluate the growth potential of E. coli O157:H7 in lettuce leaf extracts and on lettuce leaf surface at various temperatures. The pathogen can survive and multiply in the extracts and leaf surface of lettuce. The population of E. coli O157:H7 in the lettuce extracts reached to 4.79 log CFU/mL at $37^{\circ}C$. The multiplication of pathogen in lettuce extracts initiated within 10 hours of inoculation over $15^{\circ}C$ conditions. And it can survive in the lettuce leaf extracts at $4^{\circ}C$ for 100 hours at least. And this pathogen can multiply on lettuce leaf surface and the population of pathogen on the lettuce leaf surface increased to 1.82 log CFU/g at $25^{\circ}C$. At $37^{\circ}C$, the pathogen density increased to 1.53 CFU/g within 3 days after inoculation. At all temperature, irrespective of the inoculation level, similar trends in growth of E. coli O157:H7 were observed. These results emphasize the growth potential of E. coli O157:H7 in lettuce leaf extract and on lettuce leaf surface. To reduce the risk of outbreak, it is important to maintain the cold chain system during storage before the consumption.

• Archives of Pharmacal Research
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• v.21 no.3
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• pp.291-297
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• 1998

Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCI buffer (pH 7.5) containing 100 mM NaCl. Cop protein Was calculated to contain $39.1% {\alpha}-helix, 16.8% {\beta}-sheet$, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli Was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N-terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl-aminopeptidase in E. coli.

• Journal of Food Hygiene and Safety
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• v.26 no.4
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• pp.289-295
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• 2011

Produce, including leafy vegetables, has been implicated in several outbreaks of food illness. To evaluate microbiological safety of lettuce and it's cultivation area, a total of 147 samples were collected from lettuce farms and post harvest facility at Icheon, Gyeonggi province. The collected samples were assessed for presence of sanitary indicator microorganisms (Aerobic plate count, coliform count, Escherichia coli) and foodborne pathogens (Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus). The population of APC was over 4.0 log CFU from most of the samples. While the numbers of APC, and coliform of lettuce at 62 days after transplanting were 4.18 log CFU/g, and 1.00 log CFU/g, respectively, those of 10 days after transplanting were 5.37 log CFU/g, and 2.87 log CFU/g, respectively. B. cereus was highly detected from soil and balance which were contaminated with 3.5 log CFU/g, and 2.6 log CFU/100 $cm^2$, respectively. The number of E. coli recovered from gloves was 3.5 log CFU/hand. However, E. coli O157:H7, Salmonella spp., and L. monocytogenes were not detected. These data suggested that risk management system should be introduced to lettuce farms to enhance safety of lettuce.

• Journal of Microbiology and Biotechnology
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• v.14 no.6
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• pp.1327-1332
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• 2004

The hemA gene encoding 5-aminolevulinic acid synthase (ALAS) was cloned from Rhodobacter capsulatus, and its nucleotide sequence was determined. DNA sequencing data revealed one open reading frame coding for a protein with 401 amino acids that displayed high similarity to the amino acid sequences of other known ALASs. The hemA gene was then cloned and expressed under the control of constitutive promotor in E. coli. The recombinant E. coli strain was able to accumulate 5-aminolevulinic acid to 21 mM in the liquid medium supplemented with 45 mM glycine and 120 mM succinate. In addition, a marked effect of the pH of the culture medium on ALA production was observed, and the optimum pH for culture medium was determined to be 5.8-6.3.

• Journal of Microbiology and Biotechnology
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• v.7 no.5
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• pp.299-304
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• 1997

Expression in the periplasm of Escherichia coli of cloned heavy and light chain cDNAs for Fab fragment of a murine monoclonal antibody MabB9 (${\gamma}2b$, K), specific for human plasma apolipoprotein B-100 of LDL, was studied. For the purpose, a vector for two-cistronic expression of the heavy chain cDNA, at the 5' terminus, and light chain cDNA, at the 3' terminus, was constructed using the signal sequences, pelB (for heavy chain) and ompA (for light chain) in a pET vector system. The constructed vector was transformed into E. coli BL21(DE3). The expressed heavy chain (25 kDa) and light chain (23 kDa) of the antibody molecule were detected in total cell extracts as well as in the periplasmic extracts of E. coli.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.55-55
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• 1993

Farnesyl Protein transferase(FPT)는 발암유전자 ras의 단백질 산물인 p$^{21}$의 post-translational modification의 첫 단계인 ras-farnesylation에 관여하는 효소로 본 연구에서는 정제된 FPT와 E. coli에서의 발현 system을 이용하여 FPT의 구조와 기능을 밝히고 이를 FPT 방해제의 설계에 이용하고자 한다. Bovine testis에 존재하는 FPT를 30%-50%의 Ammonium sulfate로 fractionation하고, DEAE-Sephacel, Sephacryl S-300 column을 통과시킨 후 peptide(KKCVIM) affinity column을 이용하여 순수 정제하였다. 정제된 효소의 분자량은 gel-filtration에 의해 100KDa으로 추정되었고 SDS-PAGE 결과 49KDa과 46KDa의 두 subunit로 구성되었음이 확인되었다. 효소활성에는 $Mg^{2＋}$ 와 $Zn^{2＋}$가 필수적이며 최적 pH는 7.0이었다. Yeast의 FPT의 두 subunit 유전자는 Yeast genomic DNA를 template로 사용하고 각 subunit에 specific한 합성된 primer들과 vent polymerase를 이용하여 Polymerase chain reaction을 통하여 얻었다. 두 유전자를 pBluescriptII SK＋ vector를 변형시킨 두 vector, pBSK＋4와 pBChl＋4에 재조합 시킨 후 E.coli에 transformation시켜 발현시켰다. 현재 정제된 Bovine FPT와 E. coli에서 발현된 Yeast FPT의 chemical modification과 site-directed mutagenesis를 통하여 FPT의 active site와 substrate binding site에 관한 연구를 진행시키고 있다.

• Animal cells and systems
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• v.5 no.3
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• pp.195-198
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• 2001

Hepatitis B virus (HBV) polymerase, which possesses the activities of terminal binding, DNA polymerase, reverse transcriptase and RNaseH, has been shown to accomplish viral DNA replication through a pregenomic intermediate. Because the HBV polymerase has not been purified, the expression of HBV polymerase was examined in an E. coli expression system that is under the regulation of arabinose operon. The expressed individual domain containing terminal binding protein, polymerase, or RNaseH turned out to be insoluble. The activities of those domains were not able to be recovered by denaturation and renaturation using urea or guanidine-HCI. The expressed reverse transcriptase containing the polymerase and RNaseH domains became extensively degraded, whereas the proteolysis was reduced in a Ion- mutant. These results indicate that Lon protease proteolyzes the HBV reverse transcriptase expressed in E. coli.

• Korean Journal of Microbiology
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• v.22 no.4
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• pp.249-255
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• 1984

To develop the host-vector system for industrial Coryneform bacteria that seemed to be the most suitable microorganisms for molecular breeding of genes involved in the production of amion acids, nucleotides, and other products of industrial interest, broad host range E. coli plasmid R 1162 DNA was transformed into Brevibacterium ammoniagenes and the plasmids pKU6 isolated from a transformant was physically characterized. All other plasmids from the transformed cells except pKU6 exsisted as multimeric forms in Brevibacterium ammoniagenes. The plasmid DNA was retransformed into Corynebacterium glutamicum with a high frequency ($1.32{\times}10^{-1}$ per cell) and maintained stably both in Brevibacterium ammoniagenes and Corynebacterium glutamicum after 100 generations of cultures with 25-30 copy number per cell. The size of both plasmid pKU6 and plasmid R1162 were the same and restriction maps by EcoR I, Ava I, Pst I, Pvu II and Hinc II were also similar.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.217.2-217.2
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• 2003

In the type II system. there are two elongation enzymes in E. coli, FabB is well-known to its ability to elongate cis-3-decenoly-ACP (C10:1) in unsaturated fatty acid synthesis, whereas FabF is important for the thermal regulation of fatty acid composition by its ability to elongate palmitoleic acid to vaccenic acid. based on their genetic mutation anaylsis. Radiochemical enzyme assay was performed using myristoyl-ACP as a substrate, which is known for general substrate of FabB and FabF. (omitted)

• Korean Journal of Food Science and Technology
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• v.37 no.2
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• pp.287-293
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• 2005

Microbial contamination levels in commercial strawberry juices were examined for sanitary indication bacteria, such as aerobic plate count (APC), coliforms, and Escherchia coli, and pathogenic bacteria such as E. coli O157:H7, Salmonella spp., Staphylococcus aureus, and Listeria monocytogenes. APC and coliform count ranged 0-5.2 and $0-2.8\;log_{10}\;CFU/(mL,\;g,\;100cm^{2},\;hand)$, respectively, and 80% strawberry juices were contaminated with E. coli and S. aureus, detected at 19%, was found in employee's hands, strawberries, and strawberry juices, whereas E. coli O157:H7, Salmonella spp, and L. monocytogenes were not detected. These results will provide microbiological information for introduction of Hazard Analysis Critical Control Point (HACCP) system in juice shops.