猪胸膜肺炎放线杆菌外膜蛋白(OMP)5′末端保守区基因的克隆、表达及其免疫活性研究     

刘思国  彭永刚  王牟平  王春来  宫强 《中国预防兽医学报》2004,26(6)

以猪胸膜肺炎放线杆菌血清7型25-4株基因组DNA为模板,PCR方法扩增外膜蛋白(OMP)5′末端保守区基因片段(OMPc),酶切及核苷酸序列分析鉴定后,与原核表达载体质粒pGEX-6P-1进行连接,构建成重组表达载体pGEX-OM-Pc,转入大肠杆菌BL21中,以IPTG进行诱导,SDS-PAGE电泳分析发现,转化了重组质粒的菌株所表达的融合蛋白相对分子量为34 kD,与实际预测相符,命名为GST-OMPc.GST亲和层析柱进行纯化,ELISA方法对纯化蛋白进行检测.结果表明:纯化蛋白GST-OMPc能够与兔抗猪胸膜肺炎放线杆菌血清7型的阳性血清反应.OMPc蛋白的成功表达为其功能的研究打下基础.  相似文献   

捕食线虫性真菌18S rDNA基因序列与种系发生关系研究     

杨莲茹  杨晓野  刘珍莲  汪明  考桂兰  刘海成  刘军 《畜牧兽医学报》2004,35(3):329-333

对捕食线虫性真菌——少孢节丛孢菌A1分离株(Arthrobotrys oligospora A1)的18S rDNA基因序列进行了研究。结果表明：其18S rDNA全基因序列为1769bp，在进化树上与同种国外分离株A．oligospora var oligospora 1最为接近，同源性为94．7％。这与二者都具有捕食性结构—菌网的特点相一致。证实捕食线虫性真菌的捕食性结构与捕食线虫性真菌种系发生有关。从少孢节丛孢菌3个分离菌株(Arthrobotrys oligospora A1、A．oligospora 1、A．oligospora2)的进化树及同源性来看。不同国家地区的丛孢菌属(Arthrobotrys)分离株确实存在差异。  相似文献   

生长肥育猪骨骼肌注射表达pGRF基因质粒的效应研究   总被引：4，自引：1，他引：4  

张勇  周安国  吴德  朱宇旌  王康宁  杨凤 《畜牧兽医学报》2004,35(4):375-381

将猪的GRF基因表达质粒注射于猪的骨骼肌后,研究其促生长效应与机理。选用始重6.3kg的44头去势长白×太湖仔猪,分6组,采用2×3因子安排的完全随机区组设计,按6～10kg、10～20kg、20～50kg、50～100kg4个阶段饲养。4个饲养阶段的饲粮低蛋白水平分别是20.70%,17.90%,15.03%,13.00%;高蛋白水平分别是23.70%,20.90%,18.02%,16.00%。pGRF基因质粒注射剂量设0mg、0.5mg、1.0mg3个水平,于试验开始与试验猪体重达60kg时共注射基因质粒2次。测定各阶段日增重,饲料消耗量,耗料增重比以及30、70、100kg时血液中GH、GRF、IGF-I的浓度。100kg时屠宰进行胴体品质测定。结果表明:饲粮蛋白水平对各阶段及全期试验猪日增重均有显著影响(P<0.05或P<0.01),对50～100kg阶段与全期日采食量和耗料增重比有显著影响(P<0.05或P<0.01)。注射pGRF基因质粒对各阶段及全期日增重均有显著影响(P<0.05),对6～10kg、10～20kg、50～100kg3阶段及全期日采食量有显著影响(P<0.05),对6～10kg阶段、50～100kg阶段及全期耗料增重比有显著影响(P<0.05)。注射pGRF基因质粒对30kg及70kg体重时猪血液中GRF、GH、IGF-I浓度均有显著影响(P<0.05)。提高饲粮蛋白水平与注射pGRF基因质粒均可明显降低超声波测膘厚及屠宰测膘厚、增大眼肌面积(P<0.05)。  相似文献   

禽副粘病毒-2型标准毒株与不同分离株HN基因的序列测定及分析     

杨爱梅  赵继勋  赵西星  张国中  卜春亚 《畜牧兽医学报》2004,35(4):424-427

提取实验室保存禽副粘病毒-2标准毒株Yucaipa株和3株分离毒株F4、F6、F8株的RNA，经RT—PCR，获得4株毒株的HN基因，同时测得F基因与HN基因之间的序列。序列分析结果表明，HN基因的ORF全长为1743 nt，编码一个由580个氨基酸组成的蛋白。运用DNAMAN软件分析，4株毒株与GenBank上已发表毒株的HN基因的核苷酸和氨基酸之间的同源性相比，同源率分别为99．89％和99．69％。分析F基因与HN基因间序列发现两基因间序列与副粘病毒科其他病毒的基因间序列相似，含有基因起始信号和终止信号，但不含3个碱基的连接序列。通过序列分析，在分子水平上进一步证实分离于同一批进口七彩文鸟的F4、F6株是抗原性存在差异的两个毒株。毒株间的血清学关系与分子水平的核苷酸序列、氨基酸序列的比较关系一致。  相似文献   

Expression of the tobacco β-1,3-glucanase gene, PR-2d, following induction of SAR with Peronospora tabacina     

Deanna L. Funnell  Christopher B. Lawrence  Jeffrey F. Pedersen  Christopher L. Schardl 《Physiological and Molecular Plant Pathology》2004,65(6):285-296

Systemic acquired resistance (SAR) is induced following inoculation of Peronospora tabacina sporangia into the stems of Nicotiana tabacum plants highly susceptible to the pathogen. Previous results have shown that accumulation of acidic β-1,3-glucanases (PR-2's) following induction of SAR by P. tabacina may contribute to resistance to P. tabacina. We showed that up-regulation of the PR-2 gene, PR-2d, following stem inoculation with P. tabacina, is associated with SAR. Studies using plants transformed with GUS constructs containing the full length promoter from PR-2d or promoter deletions, provided evidence that a previously characterized regulatory element that is involved in response to salicylic acid (SA), may be involved in regulation of PR-2d following induction of SAR with P. tabacina. This work provides evidence that regulation of PR-2 genes during P. tabacina-induced SAR may be similar to regulation of these genes during infection of N-gene tobacco by TMV or following exogenous application of SA, and provides further support for the role of SA in regulation of genes during P. tabacina-induced SAR.  相似文献   

Analysis of a Secreted Aspartic Peptidase Disruption Mutant of Glomerella cingulata     

Kim M. Plummer  Sarah J. Clark  Lana M. Ellis  Ashwini Loganathan  Taha H. Al-Samarrai  Erik H.A. Rikkerink  Patrick A. Sullivan  Matthew D. Templeton  Peter C. Farley 《European journal of plant pathology / European Foundation for Plant Pathology》2004,110(3):265-274

Peptidases have been implicated in the pathogenicity of fungi that cause disease in plants. Expression of the secreted aspartic peptidase gene (gcsap), of a Glomerella cingulata isolate pathogenic on apples, is induced during appressorium formation. To determine whether the secreted aspartic peptidase (GcSAP) is essential to pathogenicity, gcsap was disrupted using a vector containing a 637 bp fragment of genomic DNA that encodes the sequence spanning the two active site aspartic acid (Asp) residues. To ensure that the truncated gcsap gene products could not have residual peptidase activity the codons for the active site residues Asp¹¹² and Asp²⁹⁷ were both mutated to histidine residues. Both PCR and Southern analysis confirmed disruption of gcsap. Neither gcsap mRNA nor GcSAP activity was detected in the disruption mutant. Pathogenicity tests on fruit from three apple cultivars showed that GcSAP was not required for pathogenicity. The disruption mutant grew on medium containing protein as the sole source of nitrogen because G. cingulata secretes a previously undetected peptidase(s). A serine peptidase that had a pH optimum between pH 7.0 and 8.0 and a K _m of 0.25 mM for the synthetic substrate succinyl-Ala–Ala–Pro–Phe-p-nitroanilide was identified.  相似文献   

Characterization and PCR-based Typing of Xanthomonas campestris pv. vesicatoria from Peppers and Tomatoes in Serbia     

Aleksa Obradovic  Athanassios Mavridis  Klaus Rudolph  Jaap D. Janse  Momcilo Arsenijevic  Jeffrey B. Jones  Gerald V. Minsavage  Jaw-Fen Wang 《European journal of plant pathology / European Foundation for Plant Pathology》2004,110(3):285-292

During the last two decades bacterial strains associated with necrotic leaf spots of pepper and tomato fruit spots were collected in Serbia. Twenty-eight strains isolated from pepper and six from tomato were characterized. A study of their physiological and pathological characteristics, and fatty acid composition analysis revealed that all of the strains belong to Xanthomonas campestris pv. vesicatoria. Being non-amylolytic and non-pectolytic, pathogenic on pepper but not on tomato, containing lower amounts of fatty acid 15 : 0 ante–iso, the pepper strains were designated as members of the A group of X. campestris pv. vesicatoria. However, the tomato strains hydrolyzed starch and pectate, caused compatible reactions on tomato but not on pepper, had higher percent of 15 : 0 ante–iso fatty acid, and were classified into B phenotypic group and identified as X. vesicatoria. PCR primers were developed which amplified conserved DNA regions related to the hrp genes of different strains of X. campestris pv. vesicatoria associated with pepper and tomato. Restriction analysis of the PCR product resulted in different patterns and enabled grouping of the strains into four groups. When xanthomonads isolated from pepper and tomato in Serbia were analyzed, they clustered into two groups corresponding to the grouping based on their physiological and pathological characteristics. According to the reaction of pepper and tomato differential varieties, the strains from pepper belong to races P7 and P8 and tomato strains belong to the race T2. All strains were sensitive to copper and streptomycin. Advantages and disadvantages of various bacterial spot management practices are discussed.  相似文献   

Sources of resistance to septoria tritici blotch and implications for wheat breeding   总被引：2，自引：0，他引：2  

L. Chartrain ‡  P. A. Brading  J. C. Makepeace  J. K. M. Brown † 《Plant pathology》2004,53(4):454-460

Twenty-four wheat cultivars and breeding lines were screened for isolate-specific resistance to septoria tritici blotch (STB) caused by 12 isolates of Mycosphaerella graminicola. New isolate-specific resistances that could be used in wheat breeding were identified. Major sources of resistance to STB used in world breeding programmes for decades, such as Kavkaz-K4500, Veranopolis, Catbird and TE9111, have several isolate-specific resistances. This suggests that 'pyramiding' several resistance genes in one cultivar may be an effective and durable strategy for breeding for resistance to STB in wheat. Several cultivars, including Arina, Milan and Senat, had high levels of partial resistance to most isolates tested as well as isolate-specific resistances. Resistance to isolate IPO323 was common, present in all but one of the major sources of resistance tested. This suggests that resistance to IPO323 may be an indicator of varietal resistance to STB in the field.  相似文献   

透明颤菌血红蛋白基因(vgb)在大肠杆菌中的高效表达   总被引：3，自引：0，他引：3  

祁艳霞杨明明  高文祝发明  樊俊华陈玉林 《黑龙江畜牧兽医》2005,(8):7-9

利用PCR技术将透明颠菌血红蛋白基因（vgb）克隆到融合表达载体pET28a，在大肠杆菌BL21（DE3）表达。重组蛋白在30℃诱导获得可溶表达。利用Ni^2＋亲和层析对重组蛋白进行了纯化，得到重组的透明颠菌血红蛋白，蛋白呈红色。实验现象显示重组蛋白与血红索相结合。紫外光区和可见光区波长扫描分析显示：蛋白粗提物和纯化后的血红蛋白在230nm和413nm都有强吸收峰，初步表明，尽管重组蛋白比天然蛋白多36个氨基酸，重组蛋白仍然具有生理活性。诱导后4h和6h的培养液氨基乙酰丙酸含量分别达到17mg／L和21mg／L。说明重组蛋白的表达明显促进了体内heme合成途径。  相似文献   

西安荷斯坦奶牛群5个基因座位遗传多态性的PCR-RFLP分析   总被引：6，自引：0，他引：6  

张润锋  陈宏  蓝贤勇  田燚  潘传英  胡沈荣  苏利红 《畜牧兽医学报》2005,36(6):545-549

应用PCR-RFLP方法对西安荷斯坦牛的κ-en、β-lg、β-lg5′侧翼区、CSN1S2、IGFBP-3共5个基因座位进行了多态性分析。结果表明，在西安荷斯坦牛群中，没有发现携带CSN1S2^P等位基因的个体，其多态信息含量为0。κ-en基因座位呈现低度多态(PIC=0．2366)，β-lg、β-lg5′侧翼区、IGFBP-3基因座位的多态信息含量分别为0．3168、0．3689、0．4439，均呈现中度多态。κ-en、β-lg、β-lg5′侧翼区、IGFBP-3、CSNIS2基因座位的杂合度和DNA多态度分别为0．2742、0．3947、0．4879、0．4891、0和0．0255、0．0116、0．0333、0．0112、0。而且，在西安荷斯坦牛群中，κ-en、β-lg、β-lg5′侧翼区、IGFBP-3共4个基因座位均处于Hardy-Weinberg平衡状态，CSN1S2基因座位处于纯合状态。  相似文献