• Preventive Nutrition and Food Science
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• v.15 no.2
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• pp.143-146
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• 2010

Organic zinc was included in the diet of broiler chickens to examine its effect on the skin characteristics and the expression level of skin proteins. Broiler chicks (Ross$\times$Ross) were fed a corn-wheat-soybean meal basal diet, either as control or containing an additional 80 ppm of zinc proteinate for 4 weeks, and then five broilers from each treatment were selected randomly, slaughtered, and their skin characteristics were examined. There were significant increases (p<0.05) in thigh skin epidermis and dermis thickness in the chicks fed organic zinc. Collagen content in the skin of broilers was also increased by the addition of organic zinc to the diet. 2D-gel electrophoresis patterns indicated that expression levels of the three proteins, glyoxylase 1, hypothetical protein, and dispersin B were affected by zinc feeding. These results suggest that adding organic zinc to the chicken's feed may contribute to decreased skin tearing.

• Journal of Microbiology and Biotechnology
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• v.14 no.4
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• pp.789-795
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• 2004

A promoter-probe shuttle vector pSK1Cat was constructed for the isolation of transcriptional signal sequences from Corynebacterium glutamicum. Besides conferring resistance to kanamycin in Escherichia coli and C. glutamicum, the vector carried a promoterless cat gene to confer resistance to chloramphenicol upon insertion of the appropriate transcriptional signals in the multiple cloning site. By utilizing the vector, a series of transcriptionally active fragments were isolated from the genome of C. glutamicum. The clones, ranging from 200 bp to 1 kb in size, were grouped into 3 classes of strong, medium, and weak, based on the chloramphenicol acetyltransferase (CAT) activity and sensitivity to the chloramphenicol of the clone-carrying C. glutamicum cells. C. glutamicum cells carrying the $P_{19}$ clone, a representative in the strong class, were able to grow on minimal agar plates containing over $40 mg/mell$ chloramphenicol, and showed CAT activity of 10 m㏖/mgㆍmin, performing slightly better than the cells carrying $P_{tac}$ , a strong E. coli promoter. Subcloning analysis of the $P_{19}$ clone identified a 180 bp intergenic fragment ($P_{180}$), which was located upstream of a gene encoding a hypothetical membrane protein. The expression conferred by $P_{180}$ was not affected by either the kinds of carbon sources or changes in temperature. These properties make the $P_{180}$ clone useful for the deregulated expression of biosynthetic genes in C. glutamicum during amino acid fermentation.

• Microbiology and Biotechnology Letters
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• v.38 no.4
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• pp.475-480
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• 2010

Actinomycetes are Gram-positive soil bacteria and one of the most important industrial microorganisms due to superior biosynthetic capabilities of many valuable secondary metabolites as well as production of various valuable bioconversion enzymes. Among them are cytochrome P450 hydroxylase (CYP), which are hemoproteins encoded by a super family of genes, are universally distributed in most of the organisms from all biological kingdoms. Actinomycetes are a rich source of soluble CYP enzymes, which play critical roles in the bioactivation and detoxification of a wide variety of metabolite biosynthesis and xenobiotic transformation. Cyclosporin A (CyA), one of the most commonly-prescribed immunosuppressive drugs, was previously reported to be hydroxylated at the position of 4th N-methyl leucine by a rare actinomycetes called Sebekia benihana, leading to display different biological activity spectrum such as loss of immunosuppressive activities yet retaining hair growth-stimulating side effect. In order to improve this regio-selective CyA hydroxylation in S. benihana, previously-identified several secondary metabolite up-regulatory genes from Streptomyces coelicolor and S. avermitilis were heterologously overexpressed in S. benihana using an $ermE^*$ promoter-containing Streptomyces integrative expression vector. Among tested, SCO4967 encoding a conserved hypothetical protein significantly stimulated region-specific CyA hydroxylation in S. benihana, implying that some common regulatory systems functioning in both biosynthesis and bioconversion of secondary metabolite might be present in different actinomycetes species.

• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1516-1524
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• 2014

Flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) can utilize a variety of external electron acceptors and also has stricter substrate specificity than any other glucose oxidoreductases, which makes it the ideal diagnostic enzyme in the field of glucose biosensors. A gene coding for a hypothetical protein, similar to glucose oxidase and derived from Aspergillus terreus NIH2624, was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 260,000 U/l in the culture supernatant after fed-batch cultivation for 84 h. After a three-step purification protocol that included isopropanol precipitation, affinity chromatography, and a second isopropanol precipitation, recombinant FAD-GDH was purified with a recovery of 65%. This is the first time that isopropanol precipitation has been used to concentrate a fermentation supernatant and exchange buffers after affinity chromatography purification. The purified FAD-GDH exhibited a broad and diffuse band between 83 and 150 kDa. The recombinant FAD-GDH was stable across a wide pH range (3.5 to 9.0) with maximum activity at pH 7.5 and $55^{\circ}C$. In addition, it displayed very high thermal stability, with a half-life of 82 min at $60^{\circ}C$. These characteristics indicate that FAD-GDH will be useful in the field of glucose biosensors.

• Parasites, Hosts and Diseases
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• v.51 no.4
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• pp.413-419
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• 2013

The mainstay therapy against leishmaniasis is still pentavalent antimonial drugs; however, the rate of antimony resistance is increasing in endemic regions such as Iran. Understanding the molecular basis of resistance to antimonials could be helpful to improve treatment strategies. This study aimed to recognize genes involved in antimony resistance of Leishmania tropica field isolates. Sensitive and resistant L. tropica parasites were isolated from anthroponotic cutaneous leishmaniasis patients and drug susceptibility of parasites to meglumine antimoniate (Glucantime$^{(R)}$) was confirmed using in vitro assay. Then, complementary DNA-amplified fragment length polymorphism (cDNA-AFLP) and real-time reverse transcriptase-PCR (RT-PCR) approaches were utilized on mRNAs from resistant and sensitive L. tropica isolates. We identified 2 known genes, ubiquitin implicated in protein degradation and amino acid permease (AAP3) involved in arginine uptake. Also, we identified 1 gene encoding hypothetical protein. Real-time RT-PCR revealed a significant upregulation of ubiquitin (2.54-fold), and AAP3 (2.86-fold) (P<0.05) in resistant isolates compared to sensitive ones. Our results suggest that overexpression of ubiquitin and AAP3 could potentially implicated in natural antimony resistance.

• Korean Journal of Poultry Science
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• v.34 no.2
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• pp.85-90
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• 2007

The chicken liver has been involved in various biological functions including detoxification, glycogen storage and plasma protein synthesis. The aim of this study was to investigate differentially expressed genes in chicken liver in four different growing stages. Using 10 arbitrary Annealing Control Primers (ACPs), five differentially expressed genes have been identified. Based on the Basic Local Alignment Search Tool (BLAST) search results, three of them were matched with previously known genes, and the other two were matched with unknown EST sequence and a hypothetical protein, respectively. In order to confirm the expression results, quantitative real-time PCR was also performed. The high similarities between the expression data using arbitrary ACPs and quantitative real-time PCR indicate that the identified genes are the real differentially expressed genes in different growing stages. The genes identified in this study can be used as valuable biomarkers in chicken with further investigation of the functions.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.1
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• pp.57-69
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• 2001

The chemical, elemental and biochemical components of the suspended particulate matter (SPM) were investigated in order to quantify particulate organic matter (POM) and assess diet quality for suspension feeders in the southern coastal bay systems of Korea where the marine farming of the suspension feeders are most active, The intense field observation program was carried out seasonally in the three coastal bay systems of Chinhae, Gosung and Kangjin bays, The SPM was characterized as collective properties of organic carbon (POC), nitrogen (PON), phosphorus (PP) and more refined collective properties of protein (PPr), carbohydrate (PCHO) and chlorophyll a (Chl a) compound. Although the three coastal bays are regarded as phytoplankton based ecosystem, the SPM is not composed entirely with phytoplankton cells. Due to the shallow water depth, resuspension of bottom sediment contributes significantly to some of the regions. Therefore, concentration of SPM in the surface water did not co-vary with Chl a or PPr, PCHO. In general, temporal variation of POC, PON and Chl a contents in seawater were closely associated with phytoplankton biomass in the three coastal bays, However, PPr and PCHO contents in seawater were higher in Chinhae bay than in Gosung and Kangjin bays and Chl a PPr-N ratio was higher in Chinhae bay than in Kosung and Kangjin bays, since Chinhae bay is more eutrophicated than other bays. Average C : N ratios from regressions of POC and PON of SPM were 6.6, 6.6 and 5.0 in Chinhae, Gosung and Kangjin bays, respectively. SPM in Chinhae and Gosung bays appears to be made of largely phytoplankton cells and SPM in Kangjin bay appears to be contributed from the bacterial biomass due to the shallow water depth. N : P ratios from regressions of PON and PP of SPM were 10.8 and 14.7 in spring, and 18.2 and 24.6 in Chinhae and Gosung bays, respectively. With respect to the hypothetical Redfield molecule, phytoplankton appears to be limited by the lack of N and f in spring and summer, respectively, in the two bays, In Kangjin bay, N : P ratios from regressions of PON and PP of SPM were varied from 6.3 to 12.8 throughout the year. The low N : P ratio with resepct to the hypothetical Redfield molecule, phytoplankton growth appears to be limited by the lack of N-nutrients.

• Molecules and Cells
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• v.38 no.9
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• pp.781-788
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• 2015

Mutations of MYO15A are generally known to cause severe to profound hearing loss throughout all frequencies. Here, we found two novel MYO15A mutations, c.3871C>T (p.L1291F) and c.5835T>G (p.Y1945X) in an affected individual carrying congenital profound sensorineural hearing loss (SNHL) through targeted resequencing of 134 known deafness genes. The variant, p.L1291F and p.Y1945X, resided in the myosin motor and IQ2 domains, respectively. The p.L1291F variant was predicted to affect the structure of the actin-binding site from three-dimensional protein modeling, thereby interfering with the correct interaction between actin and myosin. From the literature analysis, mutations in the N-terminal domain were more frequently associated with residual hearing at low frequencies than mutations in the other regions of this gene. Therefore we suggest a hypothetical genotype-phenotype correlation whereby MYO15A mutations that affect domains other than the N-terminal domain, lead to profound SNHL throughout all frequencies and mutations that affect the N-terminal domain, result in residual hearing at low frequencies. This genotype-phenotype correlation suggests that preservation of residual hearing during auditory rehabilitation like cochlear implantation should be intended for those who carry mutations in the N-terminal domain and that individuals with mutations elsewhere in MYO15A require early cochlear implantation to timely initiate speech development.

• Journal of Life Science
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• v.17 no.8 s.88
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• pp.1082-1089
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• 2007

Serological anlysis of recombinant cDNA expression libraries (SEREX) has led to identification of several categories of new antigens recognized by the immune system of cancer patients, which are referred to as the cancer immunome. We analyzed normal testis cDNA expression libraries with serumobtained from non-small lung cancer patient and isolated 40 distinct antigen designated KP-LuT-1 through KP-LuT-40. Among these antigens 20 antigens were previously identified by SEREX analysis of other tumor types, and 20 out of 40 antigens (50%) did not match entries in Cancer Immunome Database and were considered newly identified antigens. Sequencing analysis showed that the anti-gens comprised 26 functional known proteins and 14 noble/uncharacterized gene products. Of these, the hypothetical protein KP-LuT-6 was shown tissue-restricted. RT-PCR showed it to be expressed strongly only in normal testis. In addition to normal tissues-restricted expression, KP-LuT-6 mRNA was detected in lung tumor samples(3/l0), stomach tumor samples(3/l0), and breast tumor samples(l/5), whereas not detected in colon tumor samples(O/I2). These data suggest that KP-LuT-6 is a cancer/testis (CT)-like antigen as a potential target for cancer immunotherapies.