• KSBB Journal
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• v.6 no.3
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• pp.223-230
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• 1991

Using the shuttle vector pECCGl between Escherichia coli and Corynebacterium glutamicum and C. glutamicum strain JS231 grown in the medium supplemented with penicillin-G, which inhibits the formation of cross-links in the peptidoglycan of bacterial cell wall, various parameters involved in electroporation system including resistance, electric field strength, capacitance, DNA concentration, and cell density were investigated independently and optimized for the high efficiency transformation of coryneform bacteria. Using cells grown with 0.3U/ml of penicillin-G and harvested at A600 of 0.7-0.8, transformation efficiencies of 107-l08 transformants/$\mu\textrm{g}$ of DNA with Corynebcctertum glutamicum strain JS231 and wild type ATCC13032 were achieved under conditions of 12.5kV/cm of electric field strength, 400 ohms of resistance, $25\mu$F of capacitance, 3$\times$108 cells per transformation(1.2$\times$1010 cells/ml) and 100ng of plasmid DNA per transformation.

• Asian-Australasian Journal of Animal Sciences
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• v.7 no.4
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• pp.509-512
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• 1994

High performance liquid chromatography (HPLC) analysis was applied for determination of guanine plus cytosine (G + C) contents of DNA of Butyrivibrio fibrisolvens. By values of G + C contents, a reference strain and 20 wild strains of B. fibrisolvens were classified into at least two distinct subgroups, i.e. G + C contents of 18 strains were 38-40 mol% and those of 3 strains including the reference strain were 43-45 mol%. Clear relationships were not observed between G + C contents and biological properties of 21 strains of B. Fibrisolvens.

• Fisheries and Aquatic Sciences
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• v.6 no.1
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• pp.20-26
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• 2003

A bacterial strain was isolated from seawater to screen for phytase activities. A colony had the highest activity and was identified as an Escherichia coli strain. Using primers derived from E. coli acid phosphatase appA sequence, we cloned a 1,495 bp DNA fragment connected with the pGEM-T vector. It was over-expressed under lac promoter combined with its native promoter in E. coli $DH5\alpha$. The expression of the phytase gene occurred during late exponential growth and the intracellular phytase production was 16.9 units/ml. The yield of recombinant phytate was 412-fold higher than that of wild type E. coli K13.

• Journal of Microbiology and Biotechnology
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• v.9 no.3
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• pp.352-357
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• 1999

Enterobacter sp. B54 inhibited growth of the fungus Phytophthora capsici on potato dextrose agar (PDA). Three mutants with antifungal activities (denoted M54-47, M54-113, and M54-329) which were lost or increased, through Pl::Tn5 lac mutagenesis, were used to isolate genes responsible for fungal inhibition on PDA. Two clones were selected from the partially EcoR1-digested genomic library of the wild-type strain by probing with genomic flanking sequences of each mutant. We have isolated a 20-kb EcoR1 genomic DNA fragment from this strain that contains genes involved in hyphal growth inhibition of P. capsici on PDA. Subcloning and expression analysis of the above DNA fragment identified a 8-kb region which was necessary for antifungal activities. A 8-kb HindⅢDNA fragment covers three genomic loci inserted by Tn5 lac in each mutant. This suggested that all genes which are related to antifungal activities might be clustered in simple forms of at least 5-8 kb sizes.

• Mycobiology
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• v.41 no.3
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• pp.159-163
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• 2013

A chemical mutagenesis technique was employed for development of mutant strains of Sparassis crispa targeting the shortened cultivation time and the high ${\beta}$-glucan content. The homogenized mycelial fragments of S. crispa IUM4010 strain were treated with 0.2 vol% methyl methanesulfonate, an alkylating agent, yielding 199 mutant strains. Subsequent screening in terms of growth and ${\beta}$-glucan content yielded two mutant strains, B4 and S7. Both mutants exhibited a significant increase in ${\beta}$-glucan productivity by producing 0.254 and 0.236 mg soluble ${\beta}$-glucan/mg dry cell weight for the B4 and S7 strains, respectively, whereas the wild type strain produced 0.102 mg soluble ${\beta}$-glucan/mg dry cell weight. The results demonstrate the usefulness of chemical mutagenesis for generation of mutant mushroom strains.

• Korean Journal of Veterinary Research
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• v.59 no.2
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• pp.69-74
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• 2019

Avian malaria is one of the most important general blood parasites of poultry in Southeast Asia. Plasmodium (P.) juxtanucleare causes avian malaria in wild and domestic fowl. This study aimed to identify and characterize the Plasmodium species infecting in Thai native fowl. Blood samples were collected for microscopic examination, followed by detection of the Plasmodium cox I gene by using PCR. Five of the 10 sampled fowl had the desired 588 base pair amplicons. Sequence analysis of the five amplicons indicated that the nucleotide and amino acid sequences were homologous to each other and were closely related (100% identity) to a P. juxtanucleare strain isolated in Japan (AB250415). Furthermore, the phylogenetic tree of the cox I gene showed that the P. juxtanucleare in this study were grouped together and clustered with the Japan strain. The presence of P. juxtanucleare described in this study is the first report of P. juxtanucleare in the Thai native fowl of Thailand.

• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.392-400
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• 2019

Chondroitin, the precursor of chondroitin sulfate, which is an important polysaccharide, has drawn significant attention due to its applications in many fields. In the present study, a heterologous biosynthesis pathway of chondroitin was designed in a GRAS (generally recognized as safe) strain C. glutamicum. CgkfoC and CgkfoA genes with host codon preference were synthesized and driven by promoter Ptac, which was confirmed as a strong promoter via GFPuv reporter assessment. In a lactate dehydrogenase (ldh) deficient host, intracellular chondroitin titer increased from 0.25 to 0.88 g/l compared with that in a wild-type host. Moreover, precursor enhancement via overexpressing precursor synthesizing gene ugdA further improved chondroitin titers to 1.09 g/l. Chondroitin production reached 1.91 g/l with the engineered strain C. glutamicum ${\Delta}L-CgCAU$ in a 5-L fed-batch fermentation with a single distribution $M_w$ of 186 kDa. This work provides an alternative, safe and novel means of producing chondroitin for industrial applications.

• Journal of Microbiology
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• v.45 no.6
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• pp.492-498
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• 2007

GacS and GacA proteins form a two component signal transduction system in bacteria. Here, Tn5 transposon gacS and gacA (Gac) mutants of Pseudomonas sp. KL28, an alkylphenol degrader, were isolated by selecting for smooth colonies of strain KL28. The mutants exhibited reduced ability to migrate on a solid surface. This surface motility does not require the action of flagella unlike the well-studied swarming motility of other Pseudomonas sp. The Gac mutants also showed reduced levels of biofilm and pellicle formation in liquid culture. In addition, compared to the wild type KL28 strain, these mutants were more resistant to high concentrations of m-cresol but were more sensitive to $H_2O_2$, which are characteristics that they share with an rpoS mutant. These results indicate that the Gac regulatory cascade in strain KL28 positively controls wrinkling morphology, biofilm formation, surface translocation and $H_2O_2$ resistance, which are important traits for its capacity to survive in particular niches.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.442-449
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• 2004

Improving the light utilization efficiency of photosynthetic cells in photobioreactors (PBRs) is a major topic in algal biotechnology. Accordingly, in the current study we investigated the effect and suitability of photosynthetic pigment reduction for improving light utilization efficiency. The light-harvesting complex II (LH-II) genes of Rhodobacter sphaeroides were removed to construct a mutant strain with less pigment content. The mutant strain exhibited a slower growth rate than the wild-type under a low light intensity, while the mutant grew faster under a high light intensity. In addition, the specific absorption coefficient was lower in the mutant due to its reduced pigment content, thus it seemed that light penetrated deeper into its culture broth. However, the distance (light penetration depth) from the surface of the PBR to the compensation point did not increase, due to an increase in the compensation irradiance of the mutant strain. Experimental data showed that a reduced photosynthetic pigment content, which lessened the photoinhibition under high-intensity light, helped the volumetric productivity of photosynthetic microorganisms.

• The Korean Journal of Malacology
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• v.25 no.2
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• pp.93-96
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• 2009

Unexpected mass mortality has been one of the drawbacks in front of the stable production of Japanese scallop (Patinopecten yessoensis) on the Gangwon coasts of the East Sea. The preliminary data from our routine observation revealed that the mortality appeared to be related to variation of water temperature in the farming site and the degree of the mortality was dependent on scallop strain. The present study performed to verify the preliminary findings exhibited that the mortality was closely related to daily temperature variation rather than monthly variation. Daily temperature variation was particularly damageable to the scallop during the temperature elevation period. Scallops from hatchery seeds (Chinese strain) were more tolerant against the temperature variation over those from wild seeds. The hatchery scallop gain of the temperature tolerance was probably due to their larval experience to higher temperature in the hatchery as well as their maternal genetic acclimation to upper temperature extreme of the Chinese environment which was recently found.