한국재래산양 β-lactoglobulin 유전자 5'flanking 영역의 염기서열 분석

류승희,한성욱,서길웅,상병찬 충남대학교 농업과학연구소 2001 농업과학연구 Vol.28 No.2

본 연구는 한국 재래산양의 β-lactoglobulin(β- LG) 유전자 발현조절부위의 특성을 구명하기 위하여 PCR기법으로 specific primer를 이용하여 β-LG 발현조절부위를 증폭한 후 염기서열을 분석하여 Sheep종의 β-LG유전자 발현조절부위의 염기서열상의 차이를 분석하였다. 한국 재래산양의 genomic DNA로부터 PCR기법을 이용하여 β-LG 유전자 발현조절 부위를 증폭한 결과 Sheep종에서 보고된 바와 같이 1,077bp의 단편크기로 증폭되었음을 확인 할 수 있었다, Sheep종에서 보고된 β-LG 발현조절 부위의 염기서열과 한국재래산양에서 분석된 염기서열간의 차이는 총 897개의 염기중 46개의 nucleotide에서 차이를 나타내어 94.9%의 상동성을 보였다. 특히 한국재래산양과 Sheep종간 염기서열상의 차이는 Sheep종의 T염기가 재래산양의 C염기로 치환되었거나 반대로 Sheep종의 C염기가 재래산양에서 T염기로 치환되어 있음을 확인할 수 있었다. 이상의 결과에서 β-LG유전자 발현조절부위의 염기서열은 재래산양과 Sheep 종간에 높은 상동성을 보였으나 이들 종간의 발현조절부위의 염기서열상의 차이에 대한 유전자 발현조절 관계와 전사요소는 앞으로 연구가 더 진행되어야 할 것으로 사료된다. This study was analyzed by PCR technique with specific primer in order to investigate the characteristics of β-lactoglobulin (β-LG) gene 5'flanking region in Korean native goat. This work confirmed amplified product of 1,077 bp fragments obtained from the amplification of β-LG promoter from genomic DNA using PCR in Korean native goat. The nucleotide sequence of β-LG gene 5'flanking region in Korean native goat as compared with that of sheep β-LG were different at 46 base of 897 nucleotides, and showed high homology as about 94.9% each breed. Especially we confirmed that the difference of nucleotide sequences between Korean native goat and sheep were consisted of T→C substitution and C→T substitution reversely. As a consequences, the sequences of β-LG gene 5'flanking region showed a high homology between Korean native goat and sheep. Furthermore we should be studied that relationships between the control of gene expression and nucleotide sequences of transcription factor in Korean native goat.

PCR 기법을 이용한 한국재래산양 β-casein 유전자의 특성

김지애,류승희,유성란,이준헌,서길웅,김선균,상병찬 충남대학교 농업과학연구소 2002 농업과학연구 Vol.29 No.2

본 연구는 한국 재래 산양 112두와 유산양인 Saanen종 7두의 혈액으로부터 genomic DNA를 추출하고, PCR-RFLP 방법에 의해 β-casein 유전자의 특성을 분석하여 한국재래산양의 효율적인 유전자원의 보전 및 개량을 위한 기초 자료로 제공하고자 실시하였다. 한국재래산양의 genomic DNA로부터 PCR기법을 이용하여 β-casein의 유전자좌를 증폭한 결과 각각 481bp 크기의 단편이 양호하게 증폭되었음을 확인하였다. β-casein 유전자좌의 증폭산물에 대한 Bal Ⅰ의 제한효소를 처리한 결과, β-casein AB형은 481bp, 284bp 및 197bp의 단편을, 그리고 BB형은 284BP와 197BP의 단편을 한국재래산양과 유산양인 Saanen 종에서 확인 할 수 있었다, 유전자형 빈도에 있어서는 한국재래산양에서 β-casein AB 및 BB의 빈도는 각각 6.25 및 93.75%이었고, 유산양인 Saanen 종은 각각 57.14 및 42.86%이었다. 유전자빈도에 있어서는 한국재래산양의 β-casein A 및 B의 빈도가 각각 0.031 및 0.969이었고, Saanen 종에서는 각각 0.286 및 0.714의 빈도를 보였다 한국재래산양의 β-casein 유전자의 염기서열과 이미 보고되어 있는 goat의 염기서열(GeneBank accession Number M90556)간에는 총 11개의 염기서열에 차이를 나타내어 97.71%의 상동성을 보였다, 따라서 한국재래산양의 β-casein 유전자의 다형성과 염기서열 분석에 의한 분자유전학적 특성의 규명은 한국재래산양의 유전자원의 보전 및 개량을 위한 기초 및 응용 자료로 이용될 수 있을 것으로 생각된다. This study was performed to provide the basic data for preservation and improvement of genetic resources according to finding genetic construction obtained from analysis of genetic characteristics of β-casein gene in Korean Native goat and Saanen using the PCR-RFLP. This study confirmed the amplified products of 481bp fragments obtained from the amplification of β-casein loci by PCR. The β-casein AB genotype showed 481, 284 and 197bp, and β-casein BB genotype showed 284 and 197bp fragments in Korean Native goat and Saanen. The frequencies of β-casein genotype in Korean Native goat were 6.25 and 93.75% for AA and AB and the frequencies of β-casein genotype in Saanen were 57.14 and 42.86% for AA and AB types. The frequencies of β-casein A and B alleles were 0.031 and 0.969 in Korean Native goat and the frequencies of β-casein A and B alleles are 0.286 and 0.714 in Saanen, respectively. The nucleotide sequence of β-casein gene of Korean Native goat was 97.71% higher homology with 11 nucleotide sequences difference of that of goat reported in GeneBank (M90556). Therefore, this study of molecular genetic characteristics by the analysis of genetic polymorphism and sequencing for β-casein gene should be used as basic and applying data for preservation and improvement of genetic resources in Korean Native goat breeding.

Postweaning Multisystemic Wasting Syndrome (PMWS)에 이환된 자돈에 대해 Porcine circovirus 2 Rep 유전자 검출에 관한 연구

이용훈,김지영,楚佳奇,서상희,장경수,김명철,박창식,전무형 충남대학교 형질전환복제돼지연구센터 2007 논문집 Vol. No.10

Postweaning multisystemic wasting syndrome (PMWS) is a new emerging disease in swine herds world wide, and porcine circovirus type 2 (PCV2) is known as a major cause of PMWS. In this study, the prevalence of PCV2 Rep gene was examined for the serum, nasal swabs and feces collected from 30 heads of piglets affected with PMWS and 10 heads of normal healthy piglets using polymerase chain reaction (PCR). The diagnostic efficiency of the PCR was evaluated for different types of samples. Rep gene of the isolate (PCV2 S1) was then sequenced. In the group of PMWS-affected piglets, the prevalence of PCR-positive were 70.0% in sera, 33.3% in nasal swabs and 0% in feces. In the normal healthy group, the PCR-positive rates of sera, nasal swabs and feces were 30%, 20% and 0%, respectively. Seven piglets (17.5%) out of 40 showed the simultaneously positive results in both of sera and nasal swabs. The nucleotide sequence homology of Rep gene of PCV2 S1 as compared with the reference strain of PCV2 (GenBank accesson no.AF027217) was 99.3%.

Ciprofloxacin의 돼지 호흡기 감염증 및 설사증에 대한 치료효과

전무형,김지영,채미경,서상희,김명철,박창식,김태용,윤효인 충남대학교 수의과대학 동물의과학연구소 2003 動物醫科學硏究誌 Vol.11 No.-

To evaluate the clinical efficacy of ciprofloxacin (CFX) on treatment of the pigs with respiratory diseases or diarrhea, CFX at various dosages were administered intramuscularly to the diseased pigs selected from the pig farms located in Chungnam province. One hundreds and fourteen heads of pigs with respiratory diseases were divided into five groups and CFX at 0.25, 0.5, 1.0 ( = 2.5mg CFX/Kg B.W.) and 2.0 doses were administered i.m. for 5 days. The recovery rates were found 31.8%, 73.9%, 91.3% and 92.0%, respectively, being higher than that of the untreated control group (19.0%). The mortality of the treated groups were 0%, that is lower than that of the control (9.5%). Seven kinds of bacteria were isolated from the respiratory organs of 86 pigs, and 34 (39.5%) of 86 pigs showed the complicated infection. In analyzing by the pigs infected with a single pathogen as P. multocida, A. pleuropneumonia, Staphylclcoccus spp. or Mycoplasma spp., the recovery rates were 0∼66.6% at 0.25 dose. 66.6∼100% at 0.5 dose and 100% in all of 4 pathogens at 1.0 to 2.0 doses. At 0.25 dosage the pigs infected with A. pleuropneumonia showed the lowest recovery rate. One hundreds and twenty two heads of pigs with diarrhea were divided into five groups and CFX at 0.25, 0.5, 1.0 ( = 2.5mg CFX/Kg B.W.) and 2.0 doses were administered i.m. for 5 days. The recovery rates were 29.1%, 40.0%, 80.0% and 88.0%, respectively. The mortality of the treated groups were 0-8.3%, being much lower than that of the control. Four kinds of bacteria were isolated from the fecal specimens of 94 pigs, and 84 (89.0%) of 94 pigs showed the complicated infection.

Ciprofloxacin의 돼지 호흡기 감염증 및 설사증에 대한 치료효과

전무형,김지영,채미경,서상희,김명철,박창식,김태용,윤효인 충남대학교 형질전환복제돼지연구센터 2004 논문집 Vol. No.8

To evaluate the clinical efficacy of ciprofloxacin (CFX) on treatment of the pigs with respiratory diseases or diarrhea. CFX at various dosages were administered intramuscularly to the diseased pigs selected from the pig farms located in Chungnam province. One hundreds and fourteen heads of pigs with respiratory diseases were divided into five groups and CFX at 0.25, 0.5, 1.0 (= 2.5mg CFX/Kg B.W.) and 2.0 doses were administered i.m. for 5 days. The recovery rates were found 31.8%, 73.9%, 91.3% and 92.0%, respectively, being higher than that of the untreated control group (19.0%). The mortality of the treated groups were 0%, that is lower than that of the control (9.5%). Seven kinds of bacteria were isolated from the respiratory organs of 86 pigs, and 34 (39.5%) of 86 pigs showed the complicated infection. In analyzing by the pigs infected with a single pathogen as P. multocida; A. pleuropneumonia. Staphyloccus spp. or Mymplasma spp., the recovery rates were 0∼66.6% at 0.25 dose, 66.6∼100% at 0.5 dose and 100% in all of 4 pathogens at 1.0 to 2.0 doses. At 0.25 dosage the pigs infected with A. pleuropneumonia showed the lowest recovery rate. One hundreds and twenty two heads of pigs with diarrhea were divided into five groups and CFX at 0.25, 0.5, 1.0 (= 2.5mg CFX/Kg B.W.) and 2.0 doses were administered i.m. for 5 days. The recovery rates were 29.1 %, 40.0%, 80.0% and 88.0%, respectively. The mortality of the treated groups were 0∼8.3%, being much lower than that of the control. Four kinds of bacteria were isolated from the fecal specimens of 94 pigs, and 84 (89.0%) of 94 pigs showed the complicated infection.

Newcastle Disease Virus의 F와 HN 유전자 재조합 DNA 제조 및 면역원성

김지영,초가기,박종현,서상희,박창식,김명철,전무형 충남대학교 형질전환복제돼지연구센터 2007 논문집 Vol. No.10

Postweaning Multisystemic Wasting Syndrome (PMWS)에 이환된 자돈에 대해 Porcine circovirus 2 Rep 유전자 검출에 관한 연구

이용훈,김지영,楚佳奇,서상희,장경수,김명철,박창식,전무형 충남대학교 수의과대학 동물의과학연구소 2004 動物醫科學硏究誌 Vol.12 No.-

Postweaning multisystemic wasting syndrome (PMWS) is a new emerging disease in swine herds world wide, and porcine circovirus type 2 (PCV2) is known as a major cause of PMWS. In this study, the prevalence of PCV2 Rep gene was examined for the serum, nasal swabs and feces collected from 30 heads of piglets affected with PMWS and 10 heads of normal healthy piglets using polymerase chain reaction (PCR). The diagnostic efficiency of the PCR was evaluated for different types of samples. Rep gene of the isolate (PCV2 S1) was then sequenced. In the group of PMWS-affected piglets, the prevalence of PCR-positive were 70.0% in sera, 33.3% in nasal swabs and 0% in feces. In the normal healthy group, the PCR-positive rates of sera, nasal swabs and feces were 30%, 20% and 0%, respectively. Seven piglets (17.5%) out of 40 showed the simultaneously positive results in both of sera and nasal swabs. The nucleotide sequence homology of Rep gene of PCV2 S1 as compared with the reference strain of PCV2 (GenBank accesson no.AF027217) was 99.3%.

Roxythromycin-tylosin-furaltadone 합제의 닭 유래 세균에 대한 시험관내 항균효과

김지영,채미경,윤효인,서상희,박배근,김현수,전무형 충남대학교 수의과대학 동물의과학연구소 2003 動物醫科學硏究誌 Vol.11 No.-

In vitro antimicrobial susceptibility of roxythromycin (Rx), tylosin (Tyl), erythromycin (EM) and furaltadone (Fur) was studied, either by the single type or the combinated type of Rx-Tyl-Fur (RTF) against E. co1i (30 strains), Staph. aureus (20 strains) and Mycoplasma gallisepticum (5 strains) that are the major pathogens in poultry farm. The paper disk diffusion and MIC tests were applied. In screening test by paper disk diffusion method, the single types of antibiotics showed the sensitivity against 55 strains of bacteria in order of Fur, EM, Rx and Tyl. The combinated RTF showed the higher rate of sensitivity (80∼83.3% at 64㎍/disk, 60-75% at 32㎍/disk) and wider range of growth inhibiting zone (19.6±3.7-24.8±4.7mm) than those of the single drugs. In MIC test, the sensitivity against the 55 strains appeared in order of Fur. EM, Rx and Tyl as similar to those of paper disk diffusion method. The MIC ranges of RTF were 0.125∼16㎍/ml for E. coli, 0.25∼16㎍/ml for Staph. aureus and 1-8㎍/ml for Myco. gallisepticum, showing evidently lower than those of the single types. The MIC_(50) and MIC_(90) of RTF were estimated l∼4㎍/ml and 8∼16㎍/ml, respectively, and remarkably lower than those of the single drugs.

The pathogenesis and immunity of lethal H5N1 Hong Kong influenza viruses in animals

Seo, Sang-heui 충남대학교 수의과대학 동물의과학연구소 2002 動物醫科學硏究誌 Vol.10 No.-

In 1997, avian H5N1 influenza viruses directly jumped from chickens to humans in Hong Kong bird markets, and six humans out of 18 confirmed cases died of H5N1 influenza infections by complications of multi-organ failures including lungs. The experimental infections of mice and chickens with H5N1 influenza viruses resulted in 100% mortality with systemic infections, whereas monkeys infected nth H5N1 viruses showed sever clinical sign with virus restriction to the respiratory tracts as in humans. Although the pathogenicity of H5N1 influenza viruses in humans is not completely solved, it is clear that this virus has several potential tricks to become virulent in humans. Avian influenza viruses containing multi-basic HA cleave sites are reported to be highly virulent in chickens or probably in mice. whereas whether influenza viruses containing multi-basic HA cleave sites could be virulent in humans remains answered. 1918 Spanish flu that killed around 40 millions of humans in 6 months seemed not to have multi-basic HA cleave sites. When the deadly influenza virus like H5N1 strikes humans, there is no way to protect nave humans from deaths. In a chicken model, it showed that the cross-reactive cellular immunity alone could protect chickens from the lethality of H5N1 influenza viruses. Urgently, more efforts need to be made to design an effective vaccine to protect naive humans from a pandemic influenza virus containing avian genes.

[S10-3] Protective Effects of Korean Red Ginseng on Avian Influenza Virus

Sang Heui Seo 한국식품영양과학회 2007 한국식품영양과학회 학술대회발표집 Vol.2007 No.10