• Journal of Microbiology and Biotechnology
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• v.15 no.3
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• pp.672-677
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• 2005

The standard reference collection of strains for E. coli, the ECOR collection, was analyzed by a genome-based typing method. Seventy-one ECOR strains were subjected to repetitive extragenic palindromic PCR genome fingerprinting with BOX primers (BOX-PCR). Using a similarity value of 0.8 or more after cluster analysis of BOX-PCR fingerprinting patterns to define the same genotypes, we identified 28 genotypes in the ECOR collection. Shannon's entropy-based diversity index was 3.07, and the incident-based coverage estimator indicated potentially 420 genotypes among E. coli populations. Chi-square test of goodness-of-fit showed statistically significant association between the genotypes defined by BOX-PCR fingerprinting and the groups previously defined by multi-locus enzyme electrophoresis. This study suggests that the diversification of E. coli strains in natural populations is actively ongoing, and rep-PCR fingerprinting is a convenient and reliable method to type E. coli strains for the purposes ranging from ecology to quarantine.ine.

• Molecules and Cells
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• v.25 no.3
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• pp.446-451
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• 2008

We assessed heterologous protein expression in 64 strains obtained from the Escherichia coli Reference (ECOR) collection, a collection representing diverse natural E. coli populations. A plasmid generating a glutathione S-transferase and plant carbonic anhydrase fusion protein (GST-CA) under the control of the tac promoter was introduced into the ECOR strains, and the quantity of the fusion protein was determined by SDS-PAGE. The foreign protein was generated at various levels, from very high (40 strains, high producers) to very low (six strains, low producers). Immunoblotting showed that the high producers expressed approximately 250-500 times more GST-CA protein than the low producers. The results of semi-quantitative RT-PCR showed that the low producers generated mRNA levels comparable to those of the high producers, thereby suggesting that, at least in this case, inefficient translation is a major cause of the low production. We introduced a different plasmid, which expressed a maltose binding protein and plant guanylate kinase fusion protein (MBP-GK) into the six low producers. Interestingly, five of these expressed MBP-GK at very high levels. Thus, we conclude that the production of a particular protein from an expression vector can vary considerably, depending on the host strain. Strains in the ECOR collection could function as useful alternative hosts when a desired level of protein expression is not obtained from commonly used strains, such as E. coli K12 or B derivatives.

• Journal of the Korean Society of Marine Environment & Safety
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• v.19 no.3
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• pp.285-289
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• 2013

The coastal area of Republic of Korea is very clean compared to other countries. In this reason, west coastal area of our country is a good place for breeding up a fish such as shrimp. In winter season, the heating system is required for preventing shrimp death caused by freezing in the farm. The heater in the heating system for fishery's farm is operated very severe combating corrosion due to high accumulation by feeding material and high temperature in heated sea water. Almost all manufactured heaters of STS 316L and Ti material are scrapped every year due to heavy corrosion such a general and crevice corrosion. For comparing the general and galvanic corrosion in new heater material, the test material of Zirconium (Zr), Titanium (Ti) and STS 316L are tested by potentiodynamic polarization, electrochemical impedance spectroscopy (EIS), current density-time methods and microscopic examination in a 3.5% NaCl solution. The corrosion potential (Ecor) measured by potentiodynamic polarization for Zr, Ti and STS 316L reveals -198, -250 and -450mV, corrosion current density 0.5, 2.5 and $6.5{\mu}A/cm^2$ respectively. The film resistance measured by EIS are Zr 63,000, Ti 39,700 and 316L $3,150{\Omega}$, and the current of Zr-Ti couple is $0.03{\mu}A$, whereas Zr-316L SS is $0.1{\mu}A$. According to the result of this experiment in 3.5% NaCl solution, Zr is excellent corrosion resistance material than Ti and STS 316L.

• Microbiology and Biotechnology Letters
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• v.29 no.1
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• pp.18-24
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• 2001

Expression of the baculovirus major envelope glycoprotein gene(gp64) is regulated by transcription from botha early and late promoters. To develop a transient expression vector under the control of gp64 gene promoter, the gp64 gene of Bombyx mori nucleopolyhedrovirus-K1(BmNPV-K1) was characterized. The gp64 gene was local-ized at EcoR I-Pst I 7.38-kb fragment of the BmNPV-K1 genome. The EcorR 1-Pst I 7.38-kb fragment was cloned and the nucleotide sequence of 2,277 bases including the coding region of gp64 gene was determined. Based on these results, transient expression vector using gp64 gene promoter was constructed and named as pBm64. E.coli lacZ gene was introduced onto pBm64 as a reporter gene and expressed transiently in B. mori 5(Bm 5) cells. The expression vector transfected into the cells was maintained stably for 1 to 5 days. In order to confirm the expression of the reporter gene by gp64 promoter, recombinant virus was constructed. The recombinant virus has two independent transcription units in opposite orientations with two promoters; gp64 and polyhedrin gene promoters each initiating transcription of $\beta$-galactosidase and polyhedrin, respectively. Polyhedra formation and expression of $\beta$-galactosidase in Bm5 cells infected with the recombinant virus were observed with phase contrast microscope and in situ staining.