• Parasites, Hosts and Diseases
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• v.39 no.3
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• pp.241-246
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• 2001

The antigenic domain of the major surface protein (Nc-p43) of Neospora caninum was examined by polymerase chain reaction of its gene fragments and recombinant expression as GST fusion proteins. The fragments of Nc-p43 were as follow: a total open reading frame (OFR), T: OFR without signal sequence and C-terminal hydrophobic sequence, S: N-terminal 2/3 parts of S, A: C-terminal 2/3 parts, P; N-terminal 1/3 part, X: middle 1/3 part Y; and C-terminal 1/3 part, Z, respectively. The DNA fragments were cloned into pGEX-47 vector. Recombinant plasmids transformed into Escherichia coli of BL21 pLysS (DE3) strain were induced to express GST or GST fused fragments of Nc-p43 such as 69 kDa protein for T,66 kDa for S, 52 kDa for A,53 kDa for P, and 40 kDa proteins for X, Y, and Z, respectively in SDS-PAGE. The Nc-p43 fragments of T, S, and P reacted with a bovine serum of neosporosis while those of A, X, Y, and Z together with GST did not in the western blot. These findings suggest that the antigenic domain of Nc-p43 of N. caninum may be localized in the C-terminal 2/3 parts. Together with Al9 clone in SAGI of Toxoplasma gondii (Nam et at., 1996), the P fragment of Nc-p43 could be used as efficient antigens to diagnose and differentiate those infections with both species .

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.4
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• pp.576-583
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• 2009

G protein-coupled receptor 43 (GPR43) is a newly-discovered short-chain free fatty acid receptor and its functions remain to be defined. The objective of this study was to investigate the function of GPR43 on lipolysis. We successfully cloned the GPR43 gene from the pig (EU122439), and measured the level of GPR43 mRNA in different tissues and primary pig adipocytes. The expression level of GPR43 mRNA was higher in adipose tissue and increased gradually with adipocyte differentiation. Then we examined GPR43 mRNA level in different types, growth-stages and various regions of adipose tissue of pigs. The results showed that the expression level of GPR43 mRNA was significantly higher in adipose tissue of obese pigs than in lean pigs, and the expression level also gradually increased as age increased. We further found that the abundance of GPR43 mRNA level increased more in subcutaneous fat than visceral fat. Thereafter, we studied the correlation between GPR43 and lipid metabolism-related genes in adipose tissue and primary pig adipocytes. GPR43 gene had significant negative correlation with hormone-sensitive lipase gene (HSL, r = -0.881, p<0.01) and triacylglycerol hydrolase gene (TGH, r = -0.848, p<0.01) in adipose tissue, and had positive correlation with peroxisome proliferator-activated receptor $\gamma$ gene ($PPAR_{\gamma}$, r = 0.809, p<0.01) and lipoprotein lipase gene (LPL, r = 0.847, p<0.01) in adipocytes. In addition, we fed different concentrations of docosahexaenoic acid (DHA) to mice, and analyzed expression level changes of GPR43, HSL and TGH in adipose. The results showed that DHA down-regulated GPR43 and up-regulated HSL and TGH mRNA levels; GPR43 also had significant negative correlation with HSL (low: r = -0.762, p<0.01; high: r = -0.838, p<0.01) and TGH (low: r = -0.736, p<0.01; high: r = -0.586, p<0.01). Our results suggested that GPR43 is a potential factor which regulates lipolysis in adipose tissue, and DHA as a receptor of GPR43 might promote lipolysis through down-regulating the expression of GPR43 mRNA.

• Journal of Microbiology and Biotechnology
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• v.8 no.1
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• pp.61-66
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• 1998

A microorganism which degrades phenol and co-metabolizes trichloroethylene (TCE) was isolated from Yangsan stream after enrichment in a medium containing phenol as the sole carbon source. The isolate EL-43P was identified as the genus Rhodococcus by its morphological, cultural and physiological characteristics. Phenol-induced cells of Rhodococcus sp. EL-43P degraded TCE. Toluene and nutrient broth could not replace the phenol requirement. The optimal conditions of initial pH and temperature of media for growth were 7.0~9.0 and $30~50^{\circ}C$, respectively. Rhodococcus sp. EL-43P could grow with phenol up to 1,000 ppm. Growth was inhibited by phenol at a concentration above 1,500 ppm. It was observed that Rhodococcus sp. EL-43P was able to degrade 90% of phenol (1,000 ppm) after 40 h in a culture. Phenol-induced cells of Rhodococcus sp. EL-43P degraded 95% of $5{\mu}M$ TCE in 6 h. Rhodococcus sp. EL-43P hardly degraded TCE above $100{\mu}M$.

• KOREAN POULTRY JOURNAL
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• v.5 no.7 s.45
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• pp.43-43
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• 1973

• Parasites, Hosts and Diseases
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• v.41 no.3
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• pp.175-177
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• 2003

An ELISA was established to measure bovine IgG directed against the recombinant antigenic determinant of Nc-p43, a major surface antigen of Neospora caninum. In a previous study, two thirds of the C-terminal of the molecule was expressed as a $6{\;}{\times}{\;}His$ tagged protein (Ncp43p) for ELISA using 2/3 of the N-terminal of SAG1 from Toxoplasma gondii as a control (TgSAG1A). Among 852 cattle sera collected from stock farms scattered nation-wide, 103 sera (12.1％) were found to react with Ncp43p positively, but no positive reaction was observed with TgSAG1A. This study shows that Ncp43p could be available as an efficient antigen for the diagnosis of neosporosis in cattle. Furthermore, it together with TgSAG1A, could be useful for the differential diagnosis of N. caninum and T.gondii infections in other mammals.

• The Plant Pathology Journal
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• v.25 no.2
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• pp.136-141
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• 2009

Marking biocontrol bacteria is an essential step to monitor bacterial behavior in natural environments before application in agricultural ecosystem. In this study, we presented the simple green fluorescent protein (GFP) reporter system driven by the promoter active in Bacillus species for tagging of the biocontrol bacteria. A constitutive promoter P43 from Bacillus subtilis was fused to an enhanced promoterless gfp gene by overlap extension PCR. The GFP expression was demonstrated by the high fluorescence intensity detected in B. subtilis and Escherichia coli transformed with the P43-gfp fusion construct, respectively. The GFP reporter system was further investigated in two bacterial biocontrol strains B. licheniformis and Pseudomonas fluorescens. When the reconstructed plasmid pWH34G was introduced into B. licheniformis, GFP level measured with the fluorescence intensity in B. licheniformis was almost equivalent to that in B. subtilis. However, GFP expression level was extremely low in other biocontrol bacteria P. fluorescens by transposon based stable insertion of the P43-gfp construct into the bacterial chromosome. This study provides information regarding to the efficient biomarker P43-gfp fusion construct for bio-control Bacillus species.

• Korean Journal of Microbiology
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• v.22 no.3
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• pp.183-189
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• 1984

In order to find specific acting site of Rec A protein in plasmic polymerization in E. coli, we randomly deleted various part of pEC-3 (a derivative of pBR322) with SI nuclease treatment. Self-ligated plasmids were introduced into E. coli WA802(Rec $A^+$). A number of colonies were analyzed if they contained monomeric or polymeric plasmids by gel electrophoresis. The plasmid (pEC-43), which was deleted the region of tetracycline gene, revealed only monomeric form in Rec $A^+$ E. coli. When two plasmids, pEC-3 and pEC-43, were co-transformed in the same E. coli, the original pEC-3 showed polymerization but pEC-43 revealed monomeric form only. These results suggest that Rec A protein requires the specific site for polymerization.

• Applied Chemistry for Engineering
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• v.8 no.3
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• pp.502-509
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• 1997

Aromatic pollutants(benzene, toluene, ethylbenzene and xylenes) were photodegraded by using a UV oxidation and the rates of degradation were investigated under various reaction conditions. Each of the solution containing 50 ppm benzene, 150 ppm ethylbenzene and 250 ppm xylenes was found UV-photodegraded over 90% in 1 hour of reaction time, wheras the only was 43 % degradation was obtained with 350 ppm toluene solution. A single component solution was more degradable than a mixed component solution and benzene was almost photodegraded at a pH 4.0, 6.4 and 10.0 after reaction time is 1 hr, ehtylbenzene was photodegraded about 92%(pH 4.0), 90%(pH 6.4) and 91%(pH 10.0), xylenes was photodegraded about 95%(pH 4.0), 90%(pH 6.4) and 92%(pH 10.0), but toluene was photodegraded about 80%(pH 4.0), 43%(pH 6.4) and 70%(pH 10.0), respectively. Kinetics studies show that the rate of decay in TOC(total organic carbon) were pseudo first-order rate except ethylbenzene, and then we could evaluate mineralization rate constants(k) of aromatics.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.77-77
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• 2003

p43 is a protein with complex biological activities. It is first found as a protein associated with macromolecular tRNA synthetase complex. Within this complex, p43 specifically interacts with arginyl-tRNA synthetase to help the substrate tRNA binding to the enzyme. It is also necessary for the cellular stability of arginyl-tRNA synthetase and the molecular association of a few complex-forming tRNA synthetases. (omitted)

• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.130-139
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• 2016

A bacterial strain, producing an excellent lipolytic enzyme, was isolated from the intestinal tracts of an earthworm (Eisenia fetida). The strain was identified as Aeromonas hydrophila by phenotypic, chemotaxonomic characteristics and 16S ribosomal DNA analysis, and was designated as Aeromona hydrophila PL43. The lipolytic enzyme from A. hydrophila PL43 was purified via 35−45% ammonium sulfate precipitation, DEAE-sepharose fast flow ion-exchange, and sephacryl S-300HR gel filtration chromatography. The yield of the purified enzyme was 3.7% and 2.5% of the total activity of crude extracts with p-nitrophenyl butyrate (pNPB) and p-nitrophenyl palmitate (pNPP) as substrates, respectively. The molecular weight of the purified enzyme was approximately 74 kDa using gel filtration, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and zymography. The optimal activity of purified enzyme was observed at 50℃ and pH 8.0 using pNPB, and 60℃ and pH 8.0 using pNPP. The purified enzyme was stable in the ranges 20− 60℃ and pH 7.0−10.0. The activity of purified enzyme was inhibited by PMSF, pepstatin A, Co²⁺, Cu²⁺, and Fe²⁺, but was recovered by metal chelating of EDTA. The K_m and V_max values of the purified enzyme were 1.07 mM and 7.27 mM/min using pNPB and 1.43 mM and 2.72 mM/min using pNPP, respectively.