奶牛乳铁蛋白基因PCR-RFLP与隐性乳房炎的相关性分析

采用CMT方法检测奶牛的乳房炎发病情况,筛选120头分别设为对照组(健康奶牛)、隐性乳房炎组(试验组),每组60头,用限制性片段长度多态性(RFLP)分析技术,检测乳铁蛋白(Lf)基因启动子区域的RFLP多态性。结果表明:不同组别的奶牛Lf基因启动子区域存在RFLP多态性,说明该多态性与乳房炎存在一定关系。     

一种改进的基因表达式编程方法及应用

基因表达式编程(GEP)是基于遗传算法和遗传编程的具有更强数据处理和知识发现的进化算法。介绍了传统GEP算法的基本原理和关键技术,针对求解问题时传统GEP存在未成熟收敛和进化后期收敛速度慢等问题,提出了GEP算法的改进方法,并将改进算法应用于函数发现问题中。与传统GEP算法的对比试验表明改进的GEP算法具有更好的求解能力和更高的性能。     

洋葱AcSCL4的克隆及其功能分析

【目的】初步探讨AcSCL4在洋葱成花中的功能，为解析洋葱成花分子调控机制提供理论基础。【方法】以长日生态型洋葱品系SB2为试验材料，根据已有的洋葱转录组数据，通过qRT-PCR筛选目的基因AcSCL4后进行基因克隆，并对其进行生物信息学分析；构建AcSCL4基因的过表达载体pB221-3300-AcSCL4并转化农杆菌，采用花序浸染法浸染拟南芥，并对T₃代纯系转基因植株进行表型分析和开花相关基因表达量分析。【结果】洋葱AcSCL4基因CDS长1 515 bp，编码504个氨基酸；AcSCL4蛋白N端高度变异，C端有GRAS结构域。聚类分析发现，AcSCL4与石刁柏和棉花的SCL4蛋白亲缘关系较近。与野生型拟南芥相比，AcSCL4过表达植株表现晚花表型。荧光定量PCR结果表明，与野生型拟南芥相比,AcSCL4过表达植株开花调控基因CO和FT的表达量极显著下降。【结论】AcSCL4通过调控CO及FT基因表达来抑制洋葱开花。     

牙鲆抗菌肽hepcidin基因在成鱼组织和不同发育时期胚胎中的表达分析

抗菌肽hepcidin是由hepcidin基因编码产生的小分子多肽,是机体天然免疫的重要因子。本文利用RT-PCR的方法分析表明,抗菌肽hepcidin基因在牙鲆成鱼不同组织和不同发育时期胚胎中普遍表达,但是表达水平存在着明显的差异。     

枯草杆菌YLNF基因编码蛋白产物性质的研究

将枯草杆菌ylnF基因经PCR扩增,酶切后插入表达质粒pET15b中,转化大肠杆菌BL21(DE3),诱导表达,金属螯合亲和层析一步纯化,酶活性测定。结果表明:ylnF基因编码产物是依赖于NAD 的前咕啉-2氧化酶,一个西罗血红素生物合成末端的酶。SDS-PAGE显示YlnF亚基分子量约22kD,全酶分子量约25kD,显示酶为单体酶。将Asp102定点突变Ala102,酶活丧失,表明Asp102在催化中起重要作用。     

重庆市山羊博尔纳病病毒P24基因的检测

为了解重庆市山羊博尔纳病隐性带毒情况，分析博尔纳病病毒（BDV）的种系来源。采用巢式逆转录酶PCR结合荧光定量PCR（FQ—nRT—PCR）技术对重庆市60只山羊外周血单核细胞（PBMCs）及脑组织中的BDVP24基因片段进行了检测，将阳性产物测序，并与国外BDV毒株进行了比较。结果，山羊外周血检测阳性率为8．3％（5／60），脑组织检测阳性率为10％（6／60）。该BDVP24片段核苷酸序列与马源BDVH1766株同源性最高，达96．519／6，与标准株Strain V和He／80同源性为95．35％，并且编码的氨基酸序列也相同。表明，重庆市山羊中存在动物源性博尔纳病隐性带毒，该BDVP24核苷酸序列与Strain V和He／80株具有高度同源性。     

The role of TF-1 cell apoptosis-related gene 19 in systemic lupus erythematosus

AIM:To clarify the role of new apoptosis-related gene, TF-1 cell apoptosis-related gene 19(TFAR19), in the pathogenesis of systemic lupus erythematosis (SLE) and the relationship between TFAR19 and SLE.METHODS:DNA Ladder detection, Western blotting, immunological fluorescence method, ELISA and so on were used to test if ultraviolet B(UVB) could induce HaCaT cell apoptosis and TFAR19 expression.RESULTS:HaCaT cell apoptosis could be detected after 24 hours of 30 mj/cm² UVB irradiation. Also, we found that in active SLE patients, the TFAR19 antibody was increased, but not significant compared to the normal control.CONCLUSION:TFAR 19 is involved in the process of UVB induced ketatinocyte line HaCaT apoptosis and SLE pathogenesis.     

Cloning and Characterization of a Family of Disease Resistance Gene Analogs from 6VS of Haynaldia villosa

In the present study, microdissection of 6VS and the cloning of the resistance gene analogs (RGA) from them were reported. The 6VS were microdissected with needle and 10 types of resistance gene analogs were obtained by PCR with degenerate oligonucleotide primer designed according to resistance genes. They were designated as Hvrgak1-Hvrgak10, GenBank accession numbers are AF387113-AF387121,AY040671- AY040672. Identity among RGAs was about 10-50%, and identity with cloned R gene from plants was 5-20%. Southern hybridization analysis results showed 3 RGAs, Hvrgak2, Hvrgak4, and Hvrgak5 were linked with wheat powdery mildew resistance. These RGAs may be used as direct entrance or probes for cloning the disease resistance genes.     

Studies of Multi-Allelic Polymorphism of Dominant Dwarfing Genes in Wheat

Dwarfing breeding of wheat in the world is confined to the exploitation of recessive dwarfing sources. None of the dominant dwarfing sources discovered in common wheat (Triticum aestivum L. ) has found wide exploitation in wheat breeding due to the extreme dwarfness of their plants (20 -55 cm). We found in our work that some stable mutant lines with their plant height enhanced to different extents could be obtained in large populations derived from the stock seeds of the dominant dwarfing sources Aibian1 carrying Rht10 on 4DS and being 20 - 55 cm tall and Aisu2 carrying Rht3 on 4BS and being 55 cm tall, or from their descendants of induced mutation treatments, or from the segregating descendants of their crosses with mid- or tall-statured genotypes. Subsequently, we studied these mutation-derived lines differing in plant height with near isogenic lines and observed that the character of their enhanced plant height bred true, each carrying a semidominant dwarfing gene for a definite height and that as the plant height of the mutation-derived lines increased, the yield-contributing characters of their near isogenic lines were significantly improved. When test crosses with marker genes and physiological and biochemical genetic marker tests were performed to re-localize the semi-dominant dwarfing genes carried by the mutation-derived lines, it was confirmed that they shared common loci with Rht10 and Rht3 and that they were all mutation-derived multiple alleles. It is thus speculated that dominant dwarfing genes are of "multi-allelic polymorphism". In other words, dominant dwarfing genes, which are ultra-dwarfing, are liable to develop by mutation into a group of multiple alleles with plant height enhanced to different extents and some may have a height close to the ideal plant height for wheat breeding. Therefore, these results offer a fundamentally new approach for the exploitation of dominant dwarfing sources in wheat breeding.     

猪5-HT4b受体基因cDNA克隆和序列分析

利用RT—PCR技术克隆了猪5-HT4bR基因的核苷酸序列，并利用生物信息学手段进行了验证。核苷酸序列测定与分析表明，该基因片段全长1209bp且包含一个完整的开放阅读框，编码402个氨基酸。该序列已在GenBank登录（Accession No．AY566638）。序列分析结果表明，该基因与已报道的人、鼠等5种动物5-HT4bR基因具有较高的核酸序列和氨基酸序列同源性，分别为92．56％和93．63％。该基因编码的蛋白具有7次跨膜结构，属于G蛋白偶联受体超家族。