• International Journal of Industrial Entomology and Biomaterials
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• v.16 no.2
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• pp.87-92
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• 2008

Bombyx mori nucleopolyhedrovirus (BmNPV) orf47 gene was characterized for the first time. The coding sequence of Bm47 was amplified and subcloned into the prokaryotic expression vector pET-30a(+) in order to produce His-tagged fusion protein in the BL21 (DE3) cells. The His-Bm47 fusion protein was expressed efficiently after induction with IPTG. The purified fusion protein was used to immunize New Zealand white rabbits to prepare polyclonal antibody. As the genome of BmNPV is available in GenBank and the EST database of BmNPV is expanding, identification of novel genes of BmNPV was conceivable by data-mining techniques and bioinformatics tools. Structural bioinformatics approach to analyze the properties of Bm47 encodes protein.

• Applied Microscopy
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• v.30 no.3
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• pp.241-248
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• 2000

Electron microscopy has used for analysing the structure of protein over 30 years. Bacteriohodopsin and porins are used as examples to illustrate the progress that has recently been made in attaining resolutions which hitherto were regarded as exclusive to the realm of x-ray crystallography. To determine a protein structure used by electron microscopy, one must pass through a number of basic steps including preparation of specimen , data acquisition and data processing.

• BMB Reports
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• v.31 no.6
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• pp.585-589
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• 1998

The herpes simplex virus type-1 (HSV-1)-encoded DNA polymerase consists of two subunits, the products of the UL30 and UL42 genes. UL30 and UL42 were coexpressed in Sf9 cells infected with recombinant baculoviruses carrying the two genes. The UL30 and UL42 gene products remained tightly associated throughout the purification, which led to a near homogeneous heterodimer composed of the DNA polymerase and UL42 protein. The DNA polymerase-UL42 protein heterodimer, purified from the recombinant baculovirus-infected Sf9 cells, showed the same high degree of processivity of deoxynucleotide polymerization as the enzyme purified from the HSV-1 infected primate cells. Like the latter, it contained a 3'-5' exonuclease activity that specifically hydrolyzes an incorrectly matched nucleotide at the 3' terminus of a primer, thereby contributing to the fidelity of DNA replication.

• Korean Journal of Veterinary Research
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• v.30 no.2
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• pp.187-192
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• 1990

The Presence and quantity of protein A in Staphylococcus hyicus subsp hyicus isolates from pigs were determined by indirect hemagglutination test and enzyme-linked immunosorbent assay(ELISA). Cell-bound and extracellular protein A was demonstrated in 87.7% and 36.0% of 489 isolates, respectively, by indirect hemagglutination test. When contents of the protein A were estimated by ELISA method, all of the isolates that were positive to protein A in indirect hemagglutination test produced extracellular protein A of less than 1ng/ml. Most of the isolates produced cell-bound protein A of less than 1ng/ml, whereas 28 isolates produced 1 to 35ng/ml and 11 isolates produced 25 to 108ng/ml.

• Food Science of Animal Resources
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• v.40 no.2
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• pp.254-261
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• 2020

The aim of this study was to evaluate quality characteristics of pork myofibrillar protein (MP) added with cornstarch as affected by different pH values and salt cocnentrations. MP mixtures were prepared with three different pH values (pH 6.00, 6.25, and 6.50) and three different salt concentrations (0.15, 0.30, and 0.45 M). Cooking yield (CY), gel strength, viscosity, and scanning electron microscopy were measured to evaluate characteristics of MPs. CYs of MPs with cornstarch at above pH 6.25 or salt 0.30 M were increased compared to those at pH 6.00 or salt 0.15 M. However, gel strengths of MPs at salt 0.45 M were higher than those at salt 0.30 M. In microstructure analysis, MP gels with increasing pH value and salt concentration showed compact and uniform structure. Thus, MP gels with pH 6.25 and salt concentration of 0.30 M would be better for manufacturing meat products containing cornstarch to increase their water holding ability.

• Fisheries and Aquatic Sciences
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• v.23 no.1
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• pp.1.1-1.14
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• 2020

Background: Dietary protein requirements are dependent on a variety of factors and water temperature is one of the most important abiotic factors affecting protein requirement of fish. This study was, therefore, conducted to investigate effects of water temperature on dietary protein requirement of fry Heteropneustes fossilis which has high demand in most of the Asian markets. Methods: Quadruplicate groups of 30 fish per treatment (2.97 ± 0.65 cm; 5.11 ± 0.34 g) were fed seven isoenergetic diets (17.9 kJ g^-1 gross energy; 14.99 kJ g^-1 digestible energy) containing dietary protein levels ranging from 28 to 52% at two water temperatures (18 and 26 ℃). Experimental diets were fed to apparent satiation as semi-moist cakes thrice daily at 17:00, 12:00, and 17:30 h for 12 weeks. For precise information, various growth parameters, protein deposition, hematological parameters, metabolic enzymes, and stress response were analyzed, and effects of water temperature on dietary protein requirement was recommended on the basis of response from above parameters. Results: Groups held at 26 ℃ attained best growth, feed conversion, and protein deposition at 44% dietary protein indicating that temperature affected dietary protein requirement for optimum growth of H. fossilis fry and protein requirement seems to be satisfied with 44% dietary protein. Interestingly, interactive effects of both dietary protein levels and temperature were not found (P > 0.05). Fish reared at 18 ℃ had comparatively higher values for aspartate and alanine transferases than those reared at 26 ℃ water temperature which exhibited normal physiological value for these enzymes indicating that body metabolism was normal at this temperature. Hematological parameters also followed same pattern. Furthermore, fish reared at 26 ℃ water temperature exhibited more resistant to thermal stress (P < 0.05). The 95% maximum plateau of protein deposition data using second-degree polynomial regression analyses exhibited dietary protein requirement of fry H. fossilis between 40.8 and 41.8% of diet at 26 ℃ water temperature. The recommended range of dietary protein level and protein/digestible energy ratio for fry H. fossilis is 40.8-41.8% and 27.21-27.88 mg protein kJ^-1 digestible energy, respectively. Conclusions: Information developed is of high significance for optimizing growth potential by making better utilization of nutrient at 26 ℃ and, to develop effective management strategies for mass culture of this highly preferred fish species.

• Journal of the Korean Society of Food Science and Nutrition
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• v.2 no.1
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• pp.1-21
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• 1973

The muscle of abalone, Notohaliotis discus (REEVE), and top-shell, Turbo cornutus Solander, were examined for protein composition. Then paramyosins which are known as one of the important structural protein of the muscle fibrils were isolated from the both muscle and their physico-chemical properties such as solubility, salting-out behaviour, intrinsic viscosity, ATPase activity, etc. involving amino acid composition and N-terminal amino acid residues were investigated to elucidate phylogenie characteristics more intensively from the viewpoint of comparative biochemistry. The analysis of protein composition resulted in the following estimations: abalone muscle; water-soluble protein of 22 %, salt-soluble protein, 34%, alkali-soluble protein, 20%, and stroma protein, 24%, and top-shell muscle; water-soluble protein of 16%, salt-soluble protein, 30%, alkali-soluble protein, 29%, and stroma protein, 25%, respectively. It is demonstrated in sedimentation analysis that paramyosin and myosin-actomyosin account for approximately 65% and 35% of the salt-soluble protein of abalone, and that the composition of both sediments in top-shell was approximately 70% and 30%, respectively. The ultracentrifugally homogenous paramyosins isolated essentially according to Bailey's ethanol-dried method from both of the muscle showed a $S^{\circ}_{20,w}$ of 3. 14s for abalone and a $S^{\circ}_{20,w}$ of 3.50s for top-shell. The both paramyosins were commonly rich in arginine, aspartic acid, and glutamic acid, while scarcely contained proline and tryptophan, in rough accord with the other paramyosins thus far reported. It is clear that these gastropod paramyosins showed of having the characteristic N-terminal amino acid residues such as N-aspartic acid, N-valine, N-serine, and N-threonine in common. The abalone paramyosin completely salted in with KCl beyond $0.35{\mu}$ and the top-shell paramyosin beyond $0.30{\mu}$. The abalone paramyosin was salted-out between 18% and 30% saturation of ammonium sulphate and the top-shell paramyosin between 22% and 29% saturation. The intrinsic viscosities at abalone and top-shell paramyosins at $25^{\circ}C$ were estimated respectively to be 3.1 dl/g and 2.6 dl/g showing somewhat higher than the values for some other paramyosins from lamellibranchs. In regard with the ATPase activity, the para myosin specimens did not exhibit any significant activity over through the pH conditions of 5 to 9.5. irrespective of the presence of $Ca^{++}$ or $Mg^{++}$. So was the case with the abalone paramyosin prepared by a slightly modified Bailey's wet-extraction method.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.6
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• pp.871-880
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• 1999

The effects of dry roasting (110, 130, $150^{\circ}C$ for 15, 30, 45 min) on potential ruminant protein nutritional values in terms of: a), rumen bypass protein (BCP); b), rumen bypass starch (BST); c), fermented organic matter (FOM); d), true absorbed bypass protein (ABCP); e) microbial protein synthesized in the rumen based on available energy (E_MP); f), microbial protein synthesized in the rumen based on available nitrogen (N_MP); g), true protein supplied to the small intestine (TPSI); h), true absorbed rumen synthesized microbial protein (AMP); i), endogenous protein losses (ENDP); j), true digested protein in the small intestine (DVE); k), degraded protein balance (OEB) of whole lupin seeds (WLS) and faba beans (WFB) were evaluated by the new Dutch DV/OEB protein evaluation system. Dry roasting significantly increased BCP, BST, TPSI, ABCP, DVE (p<0.001) and decreased FOM, E_MP, AMP, N_MP and OEB (p<0.001) with increasing temperatures and times except that when temperature was at $110^{\circ}C$. The values of BCP, BST, TPSI, ABCP and DVE at $150^{\circ}C/45min$ for WLS and WFB were increased 2.2, 3.7; -, 2.0; 1.7, 1.7; 2.3, 3.7 and 1.7, 1.7 times and the values of FOM, E_MP, AMP, N_MP and OEB at $150^{\circ}C/45min$ for WLS and WFB were decreased by 15.3, 25.8; 18.1, 25.8; 18.7, 25.8; 54.6, 41.6 and 82.3% 54.7%, respectively, over the raw WLS and WFB. The results indicated that though dry roasting reduced microbial protein synthesis due to reducing FOM, TPSI didn't decrease but highly increased due to increasing BCP more than enough for compensation of the microbial protein decreasing. Therefore the net absorbable DVE in the small intestine was highly increased. The OEB values were significantly reduced for both WLS and WFB but not to the level of negative. It indicated that microbial protein synthesis might not be impaired due to the sufficient N supplied in the rumen, but the high positive OEB values in the most treatments except of $150^{\circ}C$ for 30 and 45 min of WLS (The OEB values: 54.8 and 26.0 g/kg DM) indicated that there were the large amounts of N loss in the rumen. It was concluded that dry roasting at high temperature was effective in shifting protein degradation from rumen to intestines and it increased the DVE values without reaching the negative OEB values. No optimal treatment was found in WLS due to the too high OEB values in all treatments. But dry roasting at $150^{\circ}C$ for 30 and 45 min might be optimal treatments for WLS due to the very lower OEB values.

• Parasites, Hosts and Diseases
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• v.34 no.2
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• pp.135-142
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• 1996

Antigenic domain of jai or surface protein (p30) of Toxoplosmc Sondii was analyzed after polymerase chain reaction (PCR) of its gene fragments. Hydrophilic or hydrophobic moiety of amino acid sequences were expressed as glutathione S-transferase (G57) fusion proteins. Fragments of p30 gene were as follows: 737, total p30 open reading frame (ORF) ; S28, total ORF excluding N-terminal signal sequence and C-terminal hydrophobic sequence; Al9, N-terminal 2/3 parts of A28; A19, N-terminal 2/3 of S28; P9, C-terminal 2/3 part of S28; Z9. middle 1/3 of S28; and 29, C-terminal 1/3 of S28. respectively. Primer of each fragment was synthesized to include clamp sequence of EcoR I restriction site. PCR amplified DNA was inserted info GST (26 kDa) expression vector, PGEX-47-1 to transform into Escheri,hia coei (.JM105 strain). G57 fusion proteins were expressed with IPTG induction as 63. 54, 45, 45, 35, 36. and 35 kDa proteins measured by SDS-PAGE. Each fusion protein was confirmed with G57 detection kit. Western blot analysis with the serum of a toxoplasmosis patient revealed antigenicity in proteins expressed by T37. S28, and Al9 but not those by Pl8. X9, Y10, and Z9. Antigenicity of p30 seems to be located either in N-terminal 115 part in the presence of middle 1/3 part or in the oligopeptides between margins of the first and second 1/3 parts.

• BMB Reports
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• v.30 no.3
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• pp.211-215
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• 1997

A photoactive analog of ATP, 3'(2')-O-(p-azidobenzoyl)-adenosine 5-triphosphate (AB-ATP) was synthesized by chemically coupling N-hydroxysuccinimidyl-4-azidobenzoate (NHS-AB) and ATP. The utility of AB-ATP as an effective active-site-directed photoprobe was demonstrated using catalytic subunit of protein kinase A as a model enzyme. Photoincorporation of AB-ATP was saturated with apparent dissociation constant of $30{\mu}m$ and protected completely by $100{\mu}m$ of ATP. When the enzyme was covalently modified by photolysis in the presence of saturating amounts of photoprobe, about 60% inhibition of enzyme activity was observed. These results demonstrate that AB-ATP has potential application as a probe to characterize ATP-binding proteins including protein kinases.