• Korean Journal of Agricultural Science
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• v.12 no.1
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• pp.1-16
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• 1985

This experiment was conducted to study sex expression and subsequent changes in protein and peroxidase after three cultivars of cucumber plants were treated with ethephon and gibberellin. The three cultivars of cucumber used in this study included 'Sayeup' (monoecious type), 'Sinrokdadaki' (gynoecious type), and 'Seonghowon' (intermediate type). The ethephon at 250 ppm and gibberellin at 100 ppm were treated at the 2-leaf and 4-leaf stages, and subsequent sex expression and changes in protein contents, peroxidase activities and isoperoxidase banding patterns by disc polyacrylamide gel electrophoresis were studied. The results can be summarized as follows: 1. Ethephon treatment slightly increased number of pistillate flowers and significantly decreased number of staminate flowers in the three cultivars, while gibberellin treatment significantly increased number of staminate flowers in both gynoecious 'Sinrokdadaki' and intermediate 'Seonghowon' and did not increase number of staminate flowers in monecious 'Sayeup' 2. There were some differences among three cultivars in protein contents, protein banding and isoperoxidase banding patterns of seeds and germinating seeds. However, it was not obvious to differentiate monoecious from gynoecious cultlvars by these characters. 3. Protein contents in the leaves. and stem apex after ethephon and gibberellin treatment increased gradually at the 2-leaf stage, but decreased at the 4-leaf stage. Protein contents in stem apex at the 4-leaf stage without treatment were much higher in 'Sinrokdadaki' and 'Seonghowon' than in monoecious 'Sayeup'. Protein contents in the stern apex at the 4-leaf stage were increased in the ethephon-treated monoecious 'Sayeup' and decreased in 'Sinrokdadaki' and 'Seonghowon' compared with untreated plants. 4. Peroxidase activities in the leaves and stem apex gradually decreased at the 2- leaf stage, but increased at the 4-leaf stage. Peroxidase activities in stern apex at the 4-leaf stage were significantly increased by ethephon treatment. 5. The number of protein bands in the three cuitivars after treatment gradually decreased in leaves and stem apex at the 2-leaf stage, but increased in the 4-leaf stage. The protein banding pat terns of stern apex of the ethephon-treated monoecious 'Sayeup' at the 4-leaf stage were gradually shifted to the banding.

• Korean Journal of Agricultural Science
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• v.22 no.1
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• pp.69-81
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• 1995

This study was conducted to examine the genetic variants of the blood proteins and enzymes in beef cattle breeds, Hereford, Angus and Sharolais reared at the Daekwanryuong Branch of the National Livestock Research Institute. Genetic polymorphisms of transferrin(Tf), post-transferrin2(pTf-2), albumin(Alb), post-albumin (pAlb), ceruloplasmin(Cp), amylase-I(Am-I) and hemoglobin(Hb) in blood were analyzed by the methods of PAGE(polyacrylamide gel electrophoresis) and STAGE(starch gel electrophoresis). The results obtained from this study were summarized as follows: 1. Tf and pTf-2 locus assumed to be controlled by codominant alleles, A. $D_1$, $D_2$ and E allele for Tf, F and S allele for pTf-2. In genotype frequencies, 25% and 90% for Tf $D_1D_2$ and pTf-2 SS in Hereford, 25% and 100% for Tf $AD_1$ and pTf-2 FF in Angus, 50% for Tf $D_1D_1$ and pTf-2 FS in Sharolais were found to have the highest frequency, respectively. In gene frequencies, 0.400 and 0.900 for Tf E and pTf-2 S allele in Hereford, 0.678 and 0.607 for Tf $D_1$ and pTf-2S in Sharolais were appeared to have the highest frequency. 2. Alb and pAlb locus assumed to be controlled by codominant alleles, only A allele for Alb, F and S allele for pAlb. In genotype frequencies, 70% for pAlb SS in Hereford, 90% for pAlb FF in Angus and 57.15% for pAlb SS in Sharolais were found to have the highest frequency. In gene frequencies, 0.825 and 0.750 for pAlb S in Hereford and Charolais, 0.900 for pAlb F in Angus were found to have the highest frequency. 3. Cp and Am-I locus appeared to be controlled by two alleles, F and S allele for Cp, B and C allele for Am-I. In genotype frequencies, 100% and 65% for Cp FF and Am-I BB in Hereford, 45% and 85% for Cp FF, and Am-I CC in Angus, 50% and 64.29% for Cp FF and Am-I BC in Sharolais were found to have the highest frequency. Gene frequencies were 1,000, 0.600 and 0.750 for Cp F in Herehord, Angus and Sharolais, 0.800, 0.875 and 0.680 for Am-I B, C and C allele in Hereford, Angus and sharolais, respectively. 4. Hb locus assumed to be controlled by codominant alleles, only A allele in Hereford and Angus, A and B allele in Sharolais. Genotype frequencies were 57.14% and 42.86% for Hb AA and AB in Sharolais, and gene frequencies were 0.785 and 0.215 for Hb A and B in Sharolais.

• Journal of Microbiology
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• v.44 no.5
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• pp.548-555
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• 2006

FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli (ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, $C_{83907}$ (K88ad, $CT^+,\;ST^+$). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.

• Journal of Life Science
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• v.19 no.4
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• pp.467-471
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• 2009

The entire D-loop region of the porcine mitochondrial DNA (mtDNA) was amplified from six pig breeds (Landrace, Duroc, Large White, Korean native pig, Berkshire, and Hampshire) using a primer set designed on the basis of reported porcine mtDNA sequences. From analyses through cloning, DNA sequencing and multiple sequence alignment, an 11-bp (TAAAACACTTA) duplication was observed after known tandem repetition in the D-loop region, which promoted hetroplasmy in mtDNA. Although the existence of the 11-bp duplication has been previously reported in Duroc and Japanese native pigs, there have not been any attempts to know the characteristics of this duplication in other breeds so far. A 150 bp fragment containing the 11-duplication was amplified and typed by polyacrylamide gel electrophoresis (PAGE). All Large Whites had two duplication units and Duroc showed heteromorphic patterns, 11.2% (9/80) of the animals had the 11-bp duplication in total. On the other hand, Landrace, Berkshire, Hampshire and Korean native pigs were non-duplicated. This result showed that the 11-bp duplication could be used as a breed-specific DNA marker for distinguishing pure Landrace and Large White breeds.

• Toxicological Research
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• v.13 no.1_2
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• pp.117-125
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• 1997

In order to assess the possibility whether CYP2D is involved in caffeine metabolism, we have purified and characterized the rat liver microsomal cytochrome P4502D1 (CYP2D1), equivalent to CYP2D6 in human liver, and have utilized the reconstituted CYP2D1 in the metabolism of 4 primary caffeine (1, 3, 7-trimethylxanthine) metabolites such as paraxanthine (1, 7-dimethylxanthine), 1, 3, 7-trimethylurate, theophylline (1, 3-dimethylxanthine) and theobromine (3, 7-dimethylxanthine). Rat liver CYP 2D1 has been purified to a specific content of 8.98 nmole/mg protein (13.4fold purification, 1.5% yield) using $\omega$-aminooctylagarose, hydroxlapatite, and DE52 columns in a sequential manner. As judged from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), the purified CYP2D1 was apparently homogeneous. Molecular weight of the purified CYP2D1 was found to be 51, 000 Da. Catalytic activity of the purified and then reconstituted CYP2D1 was confirmed by using bufuralol, a known subsFate of CYP2D1. The reconstituted CYP2D1 was found to produce to 1-hydroxylbufuralol at a rate of 1.43$\pm$0.13 nmol/min/nmol P450. The kinetic analysis of bufuralol hydroxylation indicated that Km and Vmax values were 7.32$\mu M$ and 1.64 nmol/min/nmol P450, respectively. The reconstituted CYP2D1 could catalyze the 7-demethylation of PX to 1-methylxanthine at a rate of 12.5 pmol/min/pmol, and also the 7- and 3- demethylations of 1, 3, 7-trimethylurate to 1, 3-dimethylurate and 1, 7-dimethylurate at 6.5 and 12.8 pmol/min/pmol CYP2D1, respectively. The reconstituted CYP2D1 could also 3-demethylate theophylline to 1-methylxanthine at 5 pmol/min/pmol and hydroxylate the theophylline to 1, 3-dimethylurate at 21.8 pmol/min/pmol CYP2D1. The reconstituted CYP2D1, however, did not metabolize TB at all (detection limits were 0.03 pmol/min/pmol). This study indicated that CYP2D1 is involved in 3-and 7-demethylations of paraxanthine and theophylline and suggested that CYP2D6 (equivalent to CYP2D1 in rat liver) present in human liver may be involved in the secondary metabolism of the primary metabolites of caffeine.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.3
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• pp.347-354
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• 2010

This study aimed to evaluate effects of 800 W microwave irradiation for 2, 4 and 6 min on chemical composition, antinutritional factors, ruminal dry matter (DM) and crude protein (CP) degradability, and in vitro CP digestibility of canola seed (CS). Nylon bags of untreated or irradiated CS were suspended in the rumen of three bulls from 0 to 48 h. Protein subfractions of untreated and microwave irradiated CS before and after incubation in the rumen were monitored by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Microwave irradiation had no effect on chemical composition of CS (p>0.05). There was a linear decrease (p<0.001) in the phytic acid and glucosinolate contents of CS as irradiation time increased. Microwave irradiation for 2, 4 and 6 min decreased the phytic acid content of CS by 8.2, 27.6 and 48.6%, respectively. The total glucosinolate contents of CS microwave irradiated for 2, 4 and 6 min decreased by 41.5, 54.7 and 59.0% respectively, compared to untreated samples. The washout fractions of DM and CP and degradation rate of the b fraction of CP decreased linearly (p<0.001) as irradiation time increased. Microwave irradiation for 2, 4 and 6 min decreased effective degradability (ED) of CP at a ruminal outflow rate of 0.05 $h^{-1}$ by 4.7, 12.3 and 21.0%, respectively. Microwave irradiation increased linearly (p<0.001) in vitro CP digestibility of ruminally undegraded CS collected after 16 h incubation. Electrophoresis results showed that napin subunits of untreated CS disappeared completely within the zero incubation period, whereas cruciferin subunits were degraded in the middle of the incubation period (16 h incubation period). In 4 and 6 min microwave irradiated CS, napin subunits were degraded after 4 and 16 h incubation periods, respectively, and cruciferin subunits were not degraded untile 24 h of incubation. In conclusion, it seems that microwave irradiation not only protected CP of CS from ruminal degradation, but also increased in vitro digestibility of CP. Moreover, microwave irradiation was effective in reducing glucosinolate and phytic acid contents of CS.

• The korean journal of orthodontics
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• v.32 no.6 s.95
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• pp.443-453
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• 2002

The principal aims of this study were to identify the composition of salivary pellicles formed on various orthodontic brackets and to obtain a detailed information about the protein adsorption profiles from whole saliva and two major glandular salivas. Four different types of orthodontic brackets were used. All were upper bicuspid brackets with a $022{\times}028$ slot Roth prescription; stainless steel metal, monocrystalline sapphire, polycrystalline alumina, and plastic brackets. Bracket pelicles were formed by the incubation of orthodontic brackets with whole saliva, submandibular-sublingual saliva, and parotid saliva for 2 hours. The bracket pellicles were extracted and confirmed by employing sodium dodecyl sulfatepolyacrylamide gel electrophoresis, Western transfer methods, and immunodetection. The results showed that low-molecular weight salivary mucin, ${\alpha}-amylase$, secretory IgA (sIgA), acidic proline-rich proteins, and cystatins were attached to all of these brackets regardless of the bracket types. High-molecular weight mucin, which promotes the adhesion of Streptococcus mutans, did not adhere to uy orthodontic brackets. Though the same components were detected in all bracket pellicles, however, the gel profiles showed qualitatively and quantitatively different pellicles, according to the origins of saliva and the bracket types. In particular, the binding of sIgA was more prominent in the pellicles from parotid saliva and the binding of cystatins was prominent in the pellicles from the form plastic brackets. This study indicates that numerous salivary proteins adhere to the orthodontic brackets and these salivary proteins adhere selectively according to bracket types and the types of the saliva.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.16
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• pp.7039-7044
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• 2015

Background: Many functional molecules controlling diverse cellular function are included in low-molecular weight proteins and peptides. Materials and Methods: To identify proteins controlling function in lung adenocarcinomas (AC), we performed two-dimensional gel electrophoresis employing tricine-SDS polyacrylamide in the second dimension (tricine 2-DE). This system was able to detect proteins under 1 kDa even with post-translational modifications. To confirm the utility of detected proteins as novel tumor markers for AC, we performed immunohistochemical analysis using 170 formalin-fixed and paraffin-embedded lung AC tissues. Results: Tricine 2-DE revealed that five proteins including S100A16 were overexpressed in lung AC-derived cells compared with lung squamous cell carcinoma, small cell carcinoma, and large cell neuroendocrine carcinoma-derived cells. Immunohistochemically, S100A16 showed various subcellular localization in lung cancer tissues and a membranous staining status was correlated with the T-factor (P=0.0008), pathological stage (P=0.0015), differentiation extent (P=0.0001), lymphatic invasion (P=0.0007), vascular invasion (P=0.0001), pleural invasion (P=0.0087), and gender (P=0.039), but not with the age or smoking history. More importantly, membranous staining of S100A16 was significantly correlated with a poorer overall survival of either stage I (P=0.0088) or stage II / III (P=0.0003) lung AC patients, and multivariate analysis confirmed that membranous expression of S100A16 was an independent adverse prognostic indicator (P=0.0001). Conclusions: The present results suggest that S100A16 protein is a novel prognostic marker for lung AC.

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

• Journal of Ginseng Research
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• v.39 no.3
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• pp.221-229
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• 2015

Background: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. Methods: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). Results: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20-O-${\beta}$-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-${\beta}$-D-Glc with the pathway $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$. However, the enzyme firstly hydrolyzed C-3 position 3-O-${\beta}$-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$, and $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$. According to enzyme kinetics, $K_m$ and $V_{max}$ of Michaelis-Menten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at $45^{\circ}C$ and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for C-Mc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. Conclusion: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPD-ginsenosides using crude enzyme.