• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.4
• /
• pp.482-486
• /
• 2007

Basic objective of this research was to compare the milk yield and composition of New Zealand White transgenic rabbit females expressing recombinant human factor VIII (hFVIII) in mammary gland during lactation with that of non-transgenic rabbit females of the same age during 30 days of lactation. Transgenic founders were generated by the microinjection of foreign DNA (mWAP-hFVIII gene construct) into the egg. F1, F2 and F3 generations of transgenic rabbits were obtained after mating of transgenic founder rabbits with non-transgenic rabbits. The amount of milk rejected was measured by weight-suckle-weight method at $10^{th}$, $20^{th}$and $30^{th}$ day of lactation. Quality of milk (content of fat, protein, lactose, dry ash, and some minerals) from transgenic and non-transgenic rabbit was also determined. Comparison of milk yield, determined by weight-suckle-weight method, showed significantly higher (p<0.05) milk production at day 20 of first lactation in non-transgenic females, but on the same day of second lactation higher milk yield was measured in transgenic ones. Significantly higher (p<0.05) content of milk fat and protein was determined in transgenic milk whilst higher content of lactose was found in non-transgenic milk. The content of minerals (calcium, phosphorus, magnesium and sodium) did not differ in both experimental and control groups. Our results showed that milk yield and composition of transgenic rabbit females (mammary specific transgenic over-expression of hFVIII) over several generations is only slightly and transiently different from milk yield of non-transgenic females, which had no significant consequence on the litter size and viability.

• Journal of Animal Science and Technology
• /
• v.45 no.3
• /
• pp.491-498
• /
• 2003

The purpose of this study is to observe purification and properties of osteopontin(OPN) from bovine milk. The purification of osteopontin from bovine milk was performed by using ion-exchange and hydrophobic chromatography. SDS-PAGE analysis revealed that the protein migrated at Mw. 60,000. NH2-terminal sequence analysis of the first seven amio acids revealed the protein to be identical to that previously reported for bovine OPN. 35-wk-old chickens, including 3 Single Comb White Leghorn (SCWL), were used to produce egg yolk antibody(IgY) against OPNas a antigen. However, the anti-OPN antibody activities determined by ELISA. Immunological assy of OPN in milk was performed using radial immunodiffusion test based on the standard curve of pure OPN. The radial precipitation lines of four different milk samples indicated that the concentrations of OPN in the milk samples were within the range of 31.7 to 39.7${\mu}g$/ml. On inhibition with OPN on precipitation of calcium phosphate, OPN was slightly higher than casein phosphopeptide(CPP) and poly-glutamic acid.

• Korean Journal of Poultry Science
• /
• v.31 no.1
• /
• pp.9-15
• /
• 2004

This study was conducted to evaluate the effects of dietary herbal plant mixture on the performance in laying hens under heat stress. One hundred ninety two 54-weeks-old ISA Brown commercial layers, were used in 56 d experimental assay. Dietary treatments included CON (control; basal diet), HPM0.05 (basal diet ＋ 0.05% herbal plant mixture), HPM0.1 (basal diet ＋ 0.1% herbal plant mixture), and HPM0.2 (basal diet ＋ 0.2% herbal plant mixture). For overall period, the hens fed with HPM0.1 and HPM0.2 diets showed lower in the hen day egg production than the hens fed with CON diet(P＜0.05). At the end of the experimental period, egg weight was heavier in HPM 0.1 treatment than in CON (P＜0.05). There were no significant differences among the treatments in egg shell breaking strength, egg shell thickness, Haugh unit, and yolk color unit. Total cholesterol concentration of yolk tended to decrease as the level of herbal plant mixture in the diet increased. Total protein of blood was higher in the hens fed with herbal plant mixture than in the hens fed with CON diet (P＜0.05). Albumin concentration of blood was increased in HPM0.05 and HPM0.1 treatments compared with CON(P＜0.05). Red blood cell (RBC) and white blood cell (WBC) concentrations in serum were increased in HPM0.1 and HPM0.2 treatments compared with CON treatment (P＜0.05). In conclusion, dietary herbal plant mixture in laying hens under heat stress adversely affected egg production but increased total protein, albumin, RBC and WBC in blood.

• Journal of Life Science
• /
• v.28 no.3
• /
• pp.300-306
• /
• 2018

Lysozymes are innate immune factors that play a critical role in the defense against pathogens in various invertebrate animals including spoon worms. In this study, an invertebrate-type lysozyme was isolated from the body wall of spoon worm, Urechis unicinctus. The acidified body wall extract was partially separated using a Sep-Pak C18 cartridge. Among the fractions, the materials that were eluted with 60% methanol/0.1% trifluoroacetic acid showed the most potent antimicrobial activity against Bacillus subtilis KCTC 1021. A series of high performance liquid chromatography (HPLC) steps were then utilized to isolate a single antimicrobial absorbance peak. The molecular weight of the antimicrobial peak was approximated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which was approximately 13 to 14 kDa. The partial primary structure of this antimicrobial protein that was analyzed, using LC-MS/MS, was CTGGRPPTCEDYAK (1611.69 Da). Homology search of these fourteen residues, using the National Center for Biotechnology Information Basic Local Alignment Search Tool (NCBI BLAST), revealed that the isolated protein was similar to the invertebrate-type lysozymes described in other animals. Then, the antimicrobial and lysozyme enzymatic (muramidase) activities of this protein were assessed. The isolated protein possessed antimicrobial activity and potent muramidase activity, which were comparable to those of hen egg white lysozyme. Therefore, the isolated protein was designated as Urechis unicinctus invertebrate-type lysozyme from the body wall, Uu-iLysb.

• Proceedings of the Korea Society of Poultry Science Conference
• /
• 2005.11a
• /
• pp.74-75
• /
• 2005

In order to evaluate dietary brown seaweed and vancomycin on the performance, layers(Isa brown) were fed on basal diet and diets containing 2.0% of brown seaweed or 10ppm vancomycin. Brown seaweed diet significantly increased(p<0.01) nitrogen balance in layer, while excretion of uric acid nitrogen and metabolizable energy utilization were not different among diets. Layer consumed more the brown seaweed diet(p<0.05). Egg production were significantly different by diets but reduced(p<0.0001) with the experimental period passed. Layer fed brown seaweed diet gave thicker shell eggs, higher Haugh unit and higher egg white CuZnSOD activity compared with those in basal diet. Also, Brown seaweed diet increased MnSOD activity in erythrocyte cytosol and peroxidase in plasma, but decreased peroxide level in plasma, and increased proliferation of PBMC stimulated with PHA-P The result indicated that brown seaweed 2.0% diet in layer improved egg quality and performance due to increased protein synthesis which were related to regulation of antioxidant system and immune cell function in blood.

• Korean Journal of Food Science and Technology
• /
• v.40 no.5
• /
• pp.568-573
• /
• 2008

The allergenicity of treated chicken egg whites (EW) was evaluated by a passive cutaneous anaphylaxis (PCA) test, immunoblot analysis, and a mouse model of food anaphylaxis. The results of the PCA test revealed that treatment with 0.3% NaOH (w/v) decreased the antigenicity of native EW to 1/4. In addition, treatment with heat ($121^{\circ}C$, 30 min) or 1% NaOH (w/v) decreased the antigenicity to 1/8 and combined treatment with 1% NaOH (w/v) and heat ($70^{\circ}C$, 15 min) decreased the antigenicity to 1/32 of that of the native EW. Immunoblot analysis revealed that the density of EW protein bands decreased in response to heat treatment, and were almost not detectable following the combined treatment. Finally, the murine model of EW anaphylaxis revealed that the mean score of systemic anaphylactic symptoms in EW challenged mice was 1.85, while the mean score in mice challenged with EW that that had been subjected to the combined treatment was only 0.20. The results of this study indicate that the most effective method of reducing EW allergenicity is combined treatment with 1% NaOH (w/v) and heat ($70^{\circ}C$, 15 min).

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.18 no.3
• /
• pp.300-306
• /
• 1989

This study was investigated to determine the functional properties of mugbean protein isolates(MPI) from sunhwa-nogdu(SH) and conventional mungbean varieties(C). MPI were prepared from defatted mungbean flour by extraction with 0.1N NaOH, precipitation at pH 4.5, washing of dispersed precipitate with buffer solution and distilled water, and subsequent freeze-drying. Crude protein content of MPI was in the range of $88.7{\sim}91.3%$. The lowest solubility was recorded at $pH\;4{\sim}5$, whereas the best buffering action was in the range of $5.5{\sim}7.5$. On the other hand, gelation of MPI was found to depend on the protein concentration. In the cases of foamability, % volume increase and specific volume were higher for 10 min. with a good whipping ability. And also, the MPI properties of two varieties of SH and C were compared and discussed.

• Korean Journal of Food Science and Technology
• /
• v.32 no.5
• /
• pp.991-996
• /
• 2000

Enzyme-linked immunosorbent assay was developed for the analysis of soy protein in foods. Competitive indirect ELISA (ciELISA) was established by using specific antibodies against the heat-stable acidic subunits (AS) of glycinin. Soy proteins in each sample used in this study were solublized in the presence of urea and DTT and boiled at $100^{\circ}C$ for 1hr and then were renatured with a cystine-containing solution. After these treatments, each isolated soy protein (ISP) heated at 60, 70, 80, $90^{\circ}C$ for 10 minutes showed almost the same curve as unheated one in the ciELISA. The detection limit of ISP was 0.3 ${\mu}g/mL$. Anti-AS antibodies have very low reactivities less than 0.1% toward non-meat proteins such as skim milk and casein and did not show any reactivities toward egg white powder and ovalbumin. When laboratory-made sausages containing ISP of $0.5{\sim}3%$ were assayed by ciELISA, the mean recovery was about 83% (C.V., 19%). In addition, the average content of soy protein in commercial sausages was 1.27%.

• Asian-Australasian Journal of Animal Sciences
• /
• v.21 no.4
• /
• pp.596-602
• /
• 2008

Ovotransferrin (OTf) was phosphorylated by dry-heating in the presence of pyrophosphate at pH 4.0 and $85^{\circ}C$ for 1 and 5 d, and the functional properties of phosphorylated OTf (PP-OTf) were investigated. The phosphorus content of OTf increased to 0.91% as a result of phosphorylation and the electrophoretic mobility of PP-OTf also increased. Although the solubility of dry-heated OTf slightly decreased, the decrease was reduced by phosphorylation. The stability against heat-induced insolubilization of OTf was somewhat improved by phosphorylation, but more than 70% of PP-OTf was insolubilized when it was heated at $70^{\circ}C$ for 10 min at pH 7.0. However, heat-induced insolubilization of PP-OTf was reduced when it was heated in the presence of phosphorylated ovalbumin. This may explain the excellent stability of phosphorylated egg white protein against heat-induced insolubilization which was reported previously. The emulsifying property of OTf was also somewhat improved by phosphorylation. The calcium phosphate-solubilizing ability of PP-OTf was enhanced. Although the degree of phosphorylation of OTf by dry-heating in the presence of pyrophosphate was similar to that of ovalbumin, the improvement of properties of PP-OTf was considerably different from those of phosphorylated ovalbumin.

• Molecules and Cells
• /
• v.44 no.7
• /
• pp.517-528
• /
• 2021

A recent genetic study with Brucella abortus revealed the secretion activator gene A (SagA) as an autolysin component creating pores in the peptidoglycan (PGN) layer for the type IV secretion system (T4SS) and peptidoglycan hydrolase inhibitor A (PhiA) as an inhibitor of SagA. In this study, we determined the crystal structures of both SagA and PhiA. Notably, the SagA structure contained a PGN fragment in a space between the N- and C-terminal domains, showing the substrate-dependent hinge motion of the domains. The purified SagA fully hydrolyzed the meso-diaminopimelic acid (DAP)-type PGN, showing a higher activity than hen egg-white lysozyme. The PhiA protein exhibiting tetrameric assembly failed to inhibit SagA activity in our experiments. Our findings provide implications for the molecular basis of the SagA-PhiA system of B. abortus. The development of inhibitors of SagA would further contribute to controlling brucellosis by attenuating the function of T4SS, the major virulence factor of Brucella.