• Clinical and Experimental Pediatrics
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• 제50권1호
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• pp.28-32
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• 2007

목 적 : ACE 유전자에 인트론 16의 287 bp 삽입(I) 혹은 결손(D)에 의한 다형성이 존재하고, 그 중 DD 유전형은 ACE 활성도가 높은 것으로 보고 되었으며 ID 다형성은 고혈압 또는 관상동맥 질환, 당뇨병성 신증, IgA 신장염 등 만성 신질환, 만성 B형 간염, 간경변증, 급성간염에서 위험인자로 알려졌다. 신생아 황달은 동아시아인이 서양인보다 2배 이상 높은 것으로 보여 유전적 연관성이 있을 것으로 사료되어 본 연구에서는 ACE 다형성과 한국인 신생아 황달과의 관계를 알아보고자 하였다. 방 법 : 혈중 빌리루빈 수치가 12 mg/dL 이상의 건강하고, 위험인자가 없는 만삭아 중 신생아 황달 환자 110명과 대조군 164명을 대상으로 하였다. 혈액을 0.5 cc를 채취하여 DNA를 분리하였고 ACE 유전자 다형성은 중합효소 연쇄반응을 이용하여 결정하였다. 1.5% agarose gel에서 전기 영동시켜 ethidium bromide로 염색한 후 유전자형을 확인하였다. 결 과 : ACE 유전자 다형성은 신생아 고빌리루빈혈증군 110명중 59명(53.6%)에서 DI 유전형을 보였고, 29명(26.4%)에서 II 유전형, 22명(20%)에서, DD 유전형을 나타냈다. 대조군 164명에서는 85명(51.8%)이 DI 유전형을 보였고, 40명(24.4%)에서 II 유전형을 보였으며, DD 유전형은 39명(23.4%)에서 나타났다. 대립유전자 빈도는 신생아 고빌리루빈혈증군에서 I 0.532, D 0.468의 분포를 보였고, 정상 대조군에서는 I 0.503, D 0.497로 비슷하였다. 결 론 : 한국 신생아에서 ACE 유전자 다형성은 DI 유전형이 많았으나, 대립유전자의 빈도는 차이가 없어 한국인 신생아 황달의 발생과 연관이 없었다.

• Asian-Australasian Journal of Animal Sciences
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• 제14권12호
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• pp.1659-1664
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• 2001

Several studies have been done regarding carcass traits and growth in pigs. Recently, these have progressed to examine increases in economic traits, including meat quality and meat quantity, by using candidate genes. One of them is the pituitary-specific protein PIT-1, a member of the POU (Pit-Oct-Unc) family of transcription factors playing an important regulatory role in developmental processes. In addition, muscle development is known to be regulated in part by growth factors and cytokines locally produced. Therefore, studies were performed to analyze PIT-1 genotypes and serum cytokines (IGF-I, IGF-II, TGF-${\beta}1$, EGF, cortisol, DHEA-S, IL-2, and IL-6) in castrated male pigs for their possible involvement in the development of carcass traits. The genotypes of PIT-1 gene were analyzed by PCR-RFLP with MspI restriction enzyme. But, only CD and DD genotypes, not CC genotype, have been detected. Based on PIT-1 genotyping, a significant difference in EGF expression beween CD type (78.8 ng/ml) and DD type (46.0 ng/ml) was detected (p<0.05), whereas other cytokines did not show any statistical significance depending on PIT-1 genotypes. Collectively, these results suggest the possibility that EGF could affect the formation of carcass traits.

• Asian-Australasian Journal of Animal Sciences
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• 제20권5호
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• pp.622-626
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• 2007

Using multi-trait animal model BLUP, selection was conducted over seven generations for growth rate (DG), real-time ultrasound loin-eye muscle area (LEA), backfat thickness (BF), and intramuscular fat content (IMF) to develop a new line of purebred Duroc pigs with enhanced meat production and meat quality. This study was intended to investigate the relationship between restriction fragment length polymorphism (RFLP) of a heart fatty acid-binding protein (H-FABP) gene and intramuscular fat content (IMF) of this Duroc purebred population. The present experiment examined the RFLP of 499 slaughtered pigs. The DNA was separated from the blood or ear tissue of the pigs, which were slaughtered at 105 kg of body weight. Intramuscular fat content of the longissimus muscle was measured using chemical analysis. A significant difference was detected in the breeding value of IMF among the H-FABP PCR RFLP genotypes. The AA genotype has a significantly larger positive effect on the IMF breeding value than do the Aa and aa genotypes for the MspI RFLP. In addition, the DD genotype has a significantly greater positive effect on IMF breeding value than the Dd and dd genotypes for the HaeIII RFLP. For the HinfI RFLP, the hh genotype has a significantly larger positive effect on IMF breeding value than the HH genotype. Multiple regression analysis was performed using the IMF breeding values as the dependent variable and the three H-FABP genotypes as independent variables. Results revealed that the contribution of the genotypes to variation in IMF breeding values was approximately 40%. These results demonstrated that H-FABP RFLPs affect IMF in this Duroc population.

• Childhood Kidney Diseases
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• 제8권1호
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• pp.18-25
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• 2004

목 적 : 알포트 증후군은 감각신경성 난청을 동반하는 만성 진행성 유전성 신질환으로, 질환의 경과의 다양성으로 인하여 진행성의 예후인자에 특히 유전학적인 역할에 대한 관심이 커지고 있다. 본 연구에서는 만성 진행성 신질환에서 신부전으로의 진행에 관여하는 것으로 알려진 안지오텐신전환효소 유전자 다형성의 발현빈도 및 병의 진행경과와의 관계를 조사하고, 신부전으로의 진행을 억제하는 치료제로서 안지오텐신전환효소 억제제의 가능성을 알아보고자 하였다. 방 법 : 임상양상 및 신조직검사상 알포트 증후군으로 진단된 12명의 환아와 발병하지 않은 12명의 가족들에서 안지오텐신전환효소 다형성의 빈도를 비교하였다. 대상 환아를 신염의 발생 후 10세 이후까지 정상적인 신기능을 유지하고 있는 신기능 유지군과, 발병 후 10세 이전에 만성신부전으로 진행을 시작한(신기능 유지기간 5년 이하) 조기 신부전 진행군으로 구분하여 신기능 감소의 진행시기와 다형성과의 관계를 비교하였고, 발병이 없는 가족에서 다형성의 빈도 및 양상을 관찰하였다. 정상 대조군으로 고혈압, 신장 혹은 심장질환이 없고, 신질환의 가족력, 안지오텐신전환효소 억제제 혹은 다른 항고혈압제의 사용경력이 없는 소아환자 35명을 정상대조군으로 선정하였다. 결과 : 1) 발병연령은 신기능 유지군이 평균 $3.45{\pm}2.4$세였고, 조기 신부전 진행군이 평균 $4.4{\pm}1.2$세로 의미있는 차이가 없었으며, 남녀비는 각각 5:3, 2:1이었다. 2) 12례의 환아 중 4례(33%)는 10세 이전에 평균 8.9세 이전에 만성신부전으로 진행하여 발병에서부터 평균 4.5년 안에 신부전으로 빠를 진행을 보여주는 조기 신부전 진행군이었고, 8례(67%)는 10세 이후까지 이상 신기능이 유지되는 신기능 유지군으로 발병 이후 평균 10.6년(최소 8년, 최대 15년 이상) 이상 신기능을 유지하고 있었으며, 두 군간의 발병 당시 신기능 및 임상양상의 의미있는 차이는 없었다. 3) 알포트 환자의 안지오텐신전환효소 유전자형태는 II type이 3례(25.0%), ID type이 5례(41.7%). DD type이 4례(33.3%)로 정상대조군의 IItype 44.3%, ID type 40.9%, DD type 14.8%와 비교하여 DD type이 많은 경향을 보였으나 두 군사이의 의미 있는 차이는 없었다. 4) 조기 신부전 진행군 4례 중 3례(75%)에서 DD type을 보였고, 신기능 유지군 8례 중 1례(12.5%)에서 DD type을 보여 두 군간의 의미있는 차이를 보였다(p<0.05). 5) 신질환을 나타내지 않은 가족을 대상으로한 조사에서는 총 12례 중 II type 5례(41.7%), ID type 5례(41.7%), DD type 2례(16.6%)로 역시 정상대조군과 유의한 차이를 보이지 않았으며, 신기능 유지여부와 가족내의 DD발현 빈도는 의미있는 관계가 없었다. 결 론 : 소아 알포트 증후군에서 안지오텐신전환효소 유전자의 다형성의 빈도는 정상대조군에 비하여 의미있는 차이가 없었으나 조기에 신부전으로 진행한 환아에서의 DD genotype의 발현율이 의미있게 높게 나타나, 예후인자로서 가능성을 시사하였으며, 따라서 안지오텐신전환효소 억제제 치료가 신부전으로의 진행을 억제하는데 도움을 줄 수 있을 것으로 생각되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권9호
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• pp.1229-1233
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• 2006

To find the candidate genes concerned with ovulation rate of sheep, Differential Display Reverse Transcription Polymerase Chain Reaction was employed to find the differently expressed cDNA controlling ovulation in the Small Tail Han sheep of polyembryony and in Tan sheep of single birth. Twenty-four primer pairs of three anchored primers and eight arbitrary primers were assembled to amplify the specialized bands from these sheep. Positive cross tests were applied to optimize the ascertainable PCR conditions in which different special bands can be identified by silver strain in one PCR tube. After eliminating the false positive PCR products by Northern hybridization, 24 differential display bands were acquired from the ovary in the Small Tail Han sheep. These EST bands were sequenced and 18 different ESTs were found in which five ESTs had several copies and 13 ESTs had only one copy. Comparing these ESTs with homologous sequences by BLAST in the GenBank, there were six ESTs with known open reading frame (ORF) and function, three ESTs with known ORF and no function, and 9 ESTs without homologous sequence. These ESTs partly represent several genes such as NOS2, tensin, TCRA, CDKN1A, ESR1 and ACTB which express especially in Small Tail Han sheep.

• BMB Reports
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• 제39권1호
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• pp.26-36
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• 2006

Differential Display PCR technique (DD-PCR) was used for the analysis of altered gene expression in hemocytes of Vibrio harveyi-infected Penaeus monodon. Forty-four combinations of arbitrary and oligo(dT) primers were used to screen for differentially expressed genes. A total of 79 differentially expressed bands could be identified from 33 primer combinations. These included 48 bands (61%) whose expression level increased and 31 bands (39%) decreased after V. harveyi challenge. Subsequently, forty-eight differential display fragments were successfully reamplified and cloned. A total of 267 clones were randomly selected and sequenced. The sequence analysis showed that 85 (31%) out of 267 clones were matched with sequences in the GenBank database which represented 24 different genes with known functions. Among the known genes, glucose transporter 1, interferon-related developmental regulator 1, lysozyme, profilin, SERPINB3, were selected for further confirmation of their differentially expression patterns by real-time PCR. The results showed increasing in expression level of the selected genes in shrimp hemocytes after microbial challenge suggesting the involvement of such genes in bacterial response in shrimp. The anti-lipopolysaccharide factor type 3 (ALFPm3) gene, previously reported in P. monodon (Supungul et al., 2002) was found among the up-regulated genes but diversity due to amino acid changes was observed. Increase in ALFPm3 transcripts upon V. harveyi injection is in accordance with that found in the previous study.

• Parasites, Hosts and Diseases
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• 제57권4호
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• pp.341-357
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• 2019

Acanthamoeba, one of free-living amoebae (FLA), remains a high risk of direct contact with this protozoan parasite which is ubiquitous in nature and man-made environment. This pathogenic FLA can cause sight-threatening amoebic keratitis (AK) and fatal granulomatous amoebic encephalitis (GAE) though these cases may not commonly be reported in our clinical settings. Acanthamoeba has been detected from different environmental sources namely; soil, water, hotspring, swimming pool, air-conditioner, or contact lens storage cases. The identification of Acanthamoeba is based on morphological appearance and molecular techniques using PCR and DNA sequencing for clinico-epidemiological purposes. Recent treatments have long been ineffective against Acanthamoeba cyst, novel anti-Acanthamoeba agents have therefore been extensively investigated. There are efforts to utilize synthetic chemicals, lead compounds from medicinal plant extracts, and animal products to combat Acanthamoeba infection. Applied nanotechnology, an advanced technology, has shown to enhance the anti-Acanthamoeba activity in the encapsulated nanoparticles leading to new therapeutic options. This review attempts to provide an overview of the available data and studies on the occurrence of pathogenic Acanthamoeba among the Association of Southeast Asian Nations (ASEAN) members with the aim of identifying some potential contributing factors such as distribution, demographic profile of the patients, possible source of the parasite, mode of transmission and treatment. Further, this review attempts to provide future direction for prevention and control of the Acanthamoeba infection.

• The Plant Pathology Journal
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• 제29권2호
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• pp.201-208
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• 2013

Drought stress is one of the major yield affecting factor for pepper plant. The effects of PGPRs were analyzed in relation with drought resistance. The PGPRs inoculated pepper plants tolerate the drought stress and survived as compared to non-inoculated pepper plants that died after 15 days of drought stress. Variations in protein and RNA accumulation patterns of inoculated and non-inoculated pepper plants subjected to drought conditions for 10 days were confirmed by two dimensional polyacrylamide gel electrophoresis (2D-PAGE) and differential display PCR (DD-PCR), respectively. A total of six differentially expressed stress proteins were identified in the treated pepper plants by 2D-PAGE. Among the stress proteins, specific genes of Cadhn, VA, sHSP and CaPR-10 showed more than a 1.5-fold expressed in amount in B. licheniformis K11-treated drought pepper compared to untreated drought pepper. The changes in proteins and gene expression patterns were attributed to the B. licheniformis K11. Accordingly, auxin and ACC deaminase producing PGPR B. licheniformis K11 could reduce drought stress in drought affected regions without the need for overusing agrochemicals and chemical fertilizer. These results will contribute to the development of a microbial agent for organic farming by PGPR.

• Molecular & Cellular Toxicology
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• 제4권4호
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• pp.293-299
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• 2008

To screen the differentially expressed genes in cadmuim-exposed marine medaka fish (Oryzias javanicus), a candidate marine test fish for ecological toxicity, the differential display polymerase chain reaction (DD-PCR) was carried out, since the genome-wide gene expression data are not available in this fish species yet. A total of 35 clones were isolated from cadmium-exposed fish and their nucleotide sequences were analyzed. The differentially expressed gene candidates were categorized to response to stimulus (3); ion binding (3); DNA binding (1); protein binding (6); carbohydrate binding (1); metabolic process (4); biological regulation (3); cellular process (2); protein synthesis (2); catalytic activity (2); sense of sight (1); immune (1); neurohormone (1); signaling activity (1); electron carrier activity (1) and others (3). For real-time quantitative RT-PCR, we selected catalase, glucose-6-phosphate dehydrogenase, heat shock protein 70, and metallothionein and confirmed that cadmium exposure enhanced induction of these four genes.

• 생명과학회지
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• 제18권8호
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• pp.1090-1095
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• 2008

근내지방 축적이 왕성하게 진행되는 12개월 및 27개월령 사이에 유전자발현이 증가하는 inducible cAMP early repressor (ICER) 유전자의 클로닝 및 유전자발현 특성을 규명하기 위하여 한우조직별, 근육 내 조지방 함량에 따라 마블링 high- 및 low 한우 각 4두와 3두씩을 공시하였다. ddRT-PCR 분석을 통하여 증폭된 300 bp PCR 산물 및 EST 서열을 중심으로 1.5 kb 한우 ICER cDNA를 염기서열 분석하였으며, 조직별 유전자발현분석결과, 식이지방이 흡수되는 소장 조직에서 ICER 유전자의 발현량이 가장 높았다. 아울러, 같은 근육타입인 등심 및 우둔조직 사이에서 ICER 유전자발현은 등심조직에서 2.5배 많이 발현하였다. 한우 마블링 high, low 두 그룹 간 유전자발현 분석 결과, high 그룹에서 ICER 유전자발현이 4배 이상 증가하는 것으로 분석되었다. 따라서, ICER 유전자는 가축의 마블링과 밀접한 관련이 있는 유전자임이 사료된다.