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嗜水气单胞菌喹诺酮类药物抗性决定区基因片段的克隆与突变分析 被引量:8
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作者 卢强 郑伟 +2 位作者 李莲瑞 刘明远 于海彬 《吉林农业大学学报》 CAS CSCD 北大核心 2003年第1期94-96,101,共4页
将分离到的4株对环丙沙星、诺氟沙星和氧氟沙星等喹诺酮类药物耐药的嗜水气单胞菌、2株敏感菌,进行旋转酶基因A亚单位(gyrA)喹诺酮抗性决定区基因(QRDR)PCR扩增。引物序列A1:AGAGTTC CTATCTTGATTACG;A2:CTGTGATGTAGGTCATCAACT,在gyrA基... 将分离到的4株对环丙沙星、诺氟沙星和氧氟沙星等喹诺酮类药物耐药的嗜水气单胞菌、2株敏感菌,进行旋转酶基因A亚单位(gyrA)喹诺酮抗性决定区基因(QRDR)PCR扩增。引物序列A1:AGAGTTC CTATCTTGATTACG;A2:CTGTGATGTAGGTCATCAACT,在gyrA基因中的位置分别为53~73和620~640。PCR扩增后,将扩增产物克隆到pMD-18T载体,酶切鉴定后测序。用DNAstar软件分析比较其蛋白序列,发现4个氨基酸突变位点,分别是83位点的Ser-Ile(4株耐药菌),92位点的Leu-Met(4株耐药菌),174位点的Ile-Phe(3株耐药菌),202、203位点的Asn、Leu-Asp、Arg(3株耐药菌,另1株Leu未突变),83位点的突变与大肠杆菌等一致,122位点具有保守的Try,耐药菌突变后的氨基酸残基分子量均比对应增大。4株耐药菌、2株敏感菌的喹诺酮抗性决定区基因片段序列已登录GenBank,注册号AY039655,AY039656,AY138539,AY138540,AY138537,AY138538。 展开更多
关键词 抗性决定性 嗜水气单胞菌 氟喹诺酮药物 GYRA基因 基因克隆 基因突变 水产养殖 病原菌
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Significance of the antigenic epitopes in heterogenous nuclear ribonucleoproteins-Ⅰfor the diagnosis and prognosis of systematic sclerosis
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作者 JING XUE ZHONG QIANG YAO +2 位作者 YANG GAO MENG XUE YU LI PING ZHU 《Journal of Microbiology and Immunology》 2006年第4期300-305,共6页
To assess the presence of autoantibodies against epitopes of heterogenous nuclear ribonucleoprotein- Ⅰ (hnRNP- Ⅰ ) in systematic sclerosis (SSc) and to analyze their clinical significance, polypeptides of hnRNP-... To assess the presence of autoantibodies against epitopes of heterogenous nuclear ribonucleoprotein- Ⅰ (hnRNP- Ⅰ ) in systematic sclerosis (SSc) and to analyze their clinical significance, polypeptides of hnRNP- Ⅰ were designed by biological technical software and analyzed with both the Wonderful Biology Information System and DNA Star-Protean Analysis Software at the same time. In these ways, two polypeptides of hnRNP- Ⅰ were obtained based on their amino acid sequences, folding features, hydrophilic, curl style, dough kneading sensation and the possibility on the surface of proteins. They are named as hnRNP- Ⅰ -1 ( NVKYNNDKSRDYTRPDLPSGDSQPSLDQT, 264-292 aa) and hnRNP- Ⅰ -2 (QLP4REGQEDQGLTKDYGNSOL, 441-461 aa), simply designated as Ⅰ -1 and Ⅰ -2. The autoantibodies against hnRNPs were detected by means of ELISA using the synthetic epitopes polypeptides as antigen. It was found that the positive rate of detection for anti- Ⅰ -1 and anti- Ⅰ -2 autoantibodies were rather higher in SSc patients than that in other CTDs and the sensitivities and specificities of the testing with ELISA for anti- Ⅰ -1 and anti- Ⅰ -2 antibodies in SSc patients were 47.62%/93.43% and 38.1%/ 91.08%, without any significant difference between these two groups of testings. Also, there was no significant difference in the clinical features and laboratory findings, such as age, involvements in digestive and respiratory tracts and erythrocyte sedimentation rate etc., between the anti- Ⅰ -Ⅰ-positive and -negative groups in SSc patients. However, the hnRNP- Ⅰ -autoantibody-positive group of patients had obviously shorter duration of disease course compared with that of the autoantibody-negative group. Anti- Ⅰ -1 and anti- Ⅰ -2 autoantibodies also had no association with antinuclear antibody, anti-Sc170 and anti-centromere antibody (ACA) in SSc patients. So, it is apparent that the autoantibodies related with SSc may act through different pathways in the pathogenesis of SSc, and the hnRNP- Ⅰ autoantibody is a new type antibody occuring during the early stage of disease and appearing to have diagnostic and prognostic significances. 展开更多
关键词 Heterogenous nuclear ribonucleoprotein- (hnRNP- Systemic sclerosis Epitope Enzyme-linked immonosorbent assay (ELISA)
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