• Preventive Nutrition and Food Science
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• 제15권2호
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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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• 제14권4호
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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.

• 한국미생물·생명공학회지
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• 제38권4호
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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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• 제24권11호
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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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• 제51권4호
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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.

• 한국가금학회지
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• 제34권2호
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• pp.85-90
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• 2007

한국 재래닭에서 성장에 따른 유전자들의 발현 변화를 알아보고 성장 촉진, 대사 및 면역 관련 유전자를 발굴하기 위하여 주령별로 닭의 간에서 RNA를 추출하였으며 10개의 arbitrary ACPs를 이용하여 차등 발현되는 유전자를 조사하였다. 발현량에 현저한 차이를 보이는 5개의 유전자들이 선별되었으며, 이 중 3개의 유전자들은 BLAST search 결과 이미 기능이 알려진 FTH1, SAA와 HSP90B1으로 밝혀졌다. 그러나 2개의 유전자들은 닭의 genome sequence가 끝났음에도 불구하고 기능이 밝혀져 있지 않아 앞으로 이 유전자들의 기능에 대한 연구가 지속되어야 함을 의미한다. 본 연구에서 닭의 간에서 성장 단계별로 발현 차이를 보이는 유전자들은 앞으로 다른 유전자와 단백질들과의 관계를 통하여 닭의 성장 및 지방 대사를 이해하는데 도움을 줄 것으로 사료된다.

• 한국수산과학회지
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• 제34권1호
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• pp.57-69
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• 2001

한반도 남해 내만역에서 계절별로 4회에 걸친 입자유기물질 농도분포를 조사한 결과 입자유기물질은 대체로 봄철과 여름철 농도가 가을과 겨울철에 비해 비교적 높다는 것을 나타내었다. 입자유기물질 농도가 높은 봄과 여름철 C : Chl a나 C . N비는 해양의 식물플랑크톤의 전형적인 비값들을 나타내고 Chl a농도 역시 이기간 중에 높게 나타나 이들 내만역에서 입자유기물질 계절 변동은 식물플랑크톤 생물량과 밀접한 상관을 갖는다는 것을 보였다. 조사 해역별 입자유기물질 성분의 농도 분포는 진해만에서 고성만과 강진만에 비해서 높은 농도를 나타내었는데, 다른 해역에 비해서 진해만에서 높은 Chl a: PPr-N비 값들은 진해만이 다른 내만역들 보다 더욱 부영양화 되었다는 것을 나타내어 부영양화 수준에 따른 식물플랑크톤 생산력이 각 내만역 사이 입자유기물질 농도의 공간적 차이를 결정하는 중요한 요인이라는 것을 시사했다. 각 내만역 내에서 입자유기물질 수평농도 분포는 소규모 내만역내에서도 공간적으로 큰 농도차를 보일 수 친다는 것을 나타내었는데, 대도시에 인접한 수역에서 높은 입자유기물질 농도와 높은 식물플랑크톤 기여는 이들 도시로부터 다량의 영양염 유입을 시사하였고, 양식장이 밀집한 수역에서 상대적으로 낮은 입자유기 물질 농도는 양식생물의 높은 섭이활동을 반영하는 것으로 고려되었다. 식물플랑크톤 성장 제한 영양염은 계절별로 각 내만역내에서 입자유기물질 C : N : P 원소 변동비로부터 평가되었는데, 진해만과 고성만에서 Redfield비와 일치하는 C : N 변동비 (6.6)는 이 해역의 입자유기물질이 식물플랑크톤에서 유래하고 있다는 것을 나타내었던 반면, 강진만에서는 5.0의 다소 낮은 C : N비를 보였다. 진해만과 고성만에서 여름철 18.2와 24.6의 높은 N : P 변동비는 인산염이 이 해역들의 식물플랑크톤 성장을 제한할 수 있는 것으로 나타나지만 봄철 10.8과 14.7의 낮은 비값은 오히려 질소계 영양염에 의한 제한을 시사한 반면, 강진만에서 N : P 변동비는 $6.3\~12.8$로 조사기간 모두 질소계 영양염이 제한 영양염으로 작용한다는 것을 시사하였다.

• Molecules and Cells
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• 제38권9호
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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.

• 생명과학회지
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• 제17권8호통권88호
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• pp.1082-1089
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• 2007

혈청학적 유전자 검색 방법(SEREX)은 암 환자의 면역계를 인식하는 종양 면역유전체(Cancer Immunome)를 형성하는 수많은 종양항원의 발견을 이끌어왔다. 본 연구는 정상인의 고환 조직으로 만들어진 cDNA liabary을 사용하여 폐암환자의 혈청으로부터 40개의 종양항원을 동정하여 그 항원들을 KP-LuT-1부터 KP-LuT-40까지 명명하였다. 이들 항원 중에서 20개는 기존의 다른 종류의 암에서 분리된 것이며 20개는 본 실험에서 새롭게 동정 된 항원들이었다. 유전자 분석을 통하여 분리된 26개의 항원들은 그 단백질의 기능이 알려진 것이었고 14개의 항원들은 기능이 분석되지 않은 유전체의 산물이었다. 이들 항원 중에서 hypothetic단백질 KP-LuT-6는 정상조직에서 제한적으로 발현되었다. RT-PCR에 의한 발현분석 결과에서 16개의 정상조직 중 고환에서만 강력하게 발현 하였고 다른 조직에서는 발현되지 않으나 폐암(3/10), 위암 (3/10) 과 유방암(1/5)들에서 발현 하였다. 이 결과는 KP-LuT-6의 항원이 암 면역치료를 위한 잠재적 유전자로 사용될 수 있는 Cancer/Testis(CT) 항원과 비슷한 유전 자로 사료된다.