梅花鹿生长激素基因的克隆及序列分析

对梅花鹿的生长激素基因I部分片段进行了克隆测序 ,序列长度为 14 6 6bp ,现已将该序列提交到GeneBank上。GeneBank中的Blast分析表明 ,梅花鹿生长激素基因I与马鹿同源性为 99% ,与鼷鹿同源性为84 %。     

鸡肠炎沙门氏菌外膜蛋白F的原核表达及免疫原性分析

根据GenBank中收录的鸡肠炎沙门氏菌OmpF基因序列设计1对引物,以鸡肠炎沙门氏菌辽宁分离株基因组为模板,扩增鸡肠炎沙门氏菌OmpF基因,用IEDB Analysis Resource在线预测分析,该基因可能存在15个线性B细胞抗原表位;同时将该基因克隆至原核表达载体PGEX-4T-1中,构建重组表达质粒PGEX-4T-1-OmpF,转入大肠杆菌BL21中,获得OmpF重组蛋白。Western-blot检测结果表明,该重组蛋白的分子质量约为66 kD(含标签蛋白),能被鸡沙门氏菌阳性血清识别,初步证实该蛋白具有免疫原性,为鸡肠炎沙门氏菌亚单位疫苗的进一步研究和开发奠定了理论基础。     

淮南猪酸肉基因的多态性分析

为了研究淮南猪的种质特性,为淮南猪保种、可持续利用及种质创新提供依据,试验采用PCR-RFLP法,对96头淮南猪的RN基因进行了检测。结果表明,RN基因在淮南猪群体中存在隐性纯合子rn^+/rn^+和杂合子RN^-/rn^+,未发现显性纯合子RN^-/RN^-。其中,基因型rn^+/rn^+和RN^-/rn^+的频率分别为0.739 6和0.260 4;等位基因rn^+和RN^-的频率分别为0.869 8和0.130 2。卡方检验表明,rn^+和RN^-基因频率处于Hardy-Weinberg平衡状态,表明群体中RN基因没有受到选择的压力。     

绵羊BMPR-IB基因作为多胎性能的候选基因在肉用多胎品种选育中的应用研究

以中国美利奴多胎细毛羊和南非肉用细毛羊为亲本，以BMPR-IB基因作为多胎性能的候选基因，进行多胎肉用品种的选育。通过对其后代多胎基因型检测分析表明：多胎这一性状的选择，符合孟德尔遗传规律，从而验证其多胎性状由一对多胎基因(FecB)控制。通过BMPR-IB基因分子标记辅助选择，多胎性状的选择仅需3个世代(3～6年)的时间就可以达到要求，对于肉用这一性状的选择，还有待于进一步选配选育。     

乳铁蛋白的生物学功能及其基因工程研究进展

乳铁蛋白是一种具有多种生物活性的糖蛋白。本文主要就乳铁蛋白的生物学功能及其基因工程研究进展作一综述。     

Laminar inflammatory gene expression in the carbohydrate overload model of equine laminitis

Reasons for performing study: There is a need to assess the laminar inflammatory response in a laminitis model that more closely resembles clinical cases of sepsis‐related laminitis than the black walnut extract (BWE) model. Objectives: To determine if a similar pattern of laminar inflammation, characterised by proinflammatory cytokine expression, occurs in the CHO model of laminitis as has been previously reported for the BWE model. Methods: Sixteen horses administered 17.6 g of starch (85% corn starch/15% wood flour)/kg bwt via nasogastric (NG) tube were anaesthetised either after developing a temperature >38.9°C (DEV group, n = 8) or at onset of Obel grade 1 lameness (OG1 group, n = 8). Control horses (CON group, n = 8) were anaesthetised 24 h after NG administration of 6 l of deionised water. Laminar tissue was collected from horses while under anaesthesia, followed by humane euthanasia. Real time‐quantitative PCR was used to assess laminar mRNA concentrations of genes involved in inflammatory signalling. Results: Increased mRNA concentrations (P<0.05) for IL‐1β, IL‐6, IL‐12p35, COX‐2, E‐selectin and ICAM‐1 were present in laminae from horses with OG1 lameness but not at the DEV time, when compared to the CON horses. No differences between the groups were found for IL‐2, IL‐4, IL‐10, TNF‐α, IFN‐γ or COX‐1 at either the DEV or OG1 time points. Conclusions: There was a notable difference in the temporal pattern of inflammatory events between the BWE and CHO models, with the majority of laminar inflammatory events appearing to occur at or near the onset of lameness in the CHO model, whereas many of these events peak earlier in the developmental stages in the BWE model. This suggests that, in addition to circulating inflammatory molecules, there may be a local phenomenon in the CHO model resulting in the simultaneous onset of multiple laminar events including endothelial activation, leucocyte emigration and proinflammatory cytokine expression. Potential relevance: The similar (although somewhat delayed) inflammatory response in the CHO model of laminitis indicates that inflammatory signalling is a consistent entity in the pathophysiology of laminitis.     

血红素氧合酶-2基因真核表达载体的构建

为构建血红素氧合酶-2（HO-2）真核表达载体pEF1α-HO-2-AcGFP,并观察其在小鼠脑血管内皮细胞中的表达情况。试验利用PCR技术从C57BL/6小鼠海马组织中扩增出HO-2基因cDNA序列,用双酶切法将此序列克隆到真核表达载体pEF1α-IRES-AcGFP上,构建HO-2的真核表达载体pEF1α-HO-2-AcGFP,重组载体经EcoRI和BamHI双酶切和测序鉴定。电穿孔法转染小鼠脑血管内皮细胞,48h后,实时定量PCR、Western blot检测HO-2在小鼠脑血管内皮细胞中mRNA和蛋白质水平的表达情况。结果表明,pEF1α-HO-2-AcGFP载体构建正确;重组载体转染小鼠脑血管内皮细胞后HO-2在mRNA水平表达量较空载体转染极显著增高（P〈0.01）,在蛋白水平表达量较空载体转染显著增高（P〈0.05）。表明HO-2基因的真核表达载体pEF1α-HO-2-AcGFP构建成功,为进一步研究其机制和功能奠定了基础。     

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。     

Determination of nucleotide sequence of SRY gene in sika deer (Cervus nippon)

The aim of the present study was to determine the whole nucleotide sequence of the open reading frame of the sex‐determining region Y (SRY‐ORF) in wild sika deer. The SRY gene of wild sika deer was obtained by polymerase chain reaction (PCR) with DNA from blood samples. The whole nucleotide sequence of the SRY‐ORF in wild sika deer consisted of 687 bp and encoded 229 deduced amino acids. In comparison with the bovine SRY gene, the percentage of nucleotide sequence homology was 91.0% in the overall ORF, and those of the N‐terminal, high mobility group (HMG) box, and C‐terminal regions within ORF were 88.9%, 96.2% and 87.9%, respectively. The nucleotide sequences of sika deer SRY‐ORF characterized in the present study can be used for phylogenetic analysis or sexing in wild sika deer.     

细胞凋亡的常用调控基因研究近况

细胞凋亡主要受基因的控制，现已发现许多基因对细胞凋亡均有直接或间接的调控作用。通常将这些基因分为两大类，即细胞生存基因和细胞死亡基因。一般认为与细胞生存有关的基因包括C－myc,C-abl,Ras,V-src,Bcl-2,EIB,LMW5-HL,C-kit和Bcl-x等；与细胞死亡有关的基因包含P^35,P^53,RB,DCC,WT-1,CpGv,Bcr-abl,V-erb-A,tats20,DAD1,ADO/Fas,TGF-β，TRP－2，RP－8，TRPM－2，SGP－2，TIA，Bax,ICE,C-rel,irrecrst等。本文对目前使用较多研究较深入的几种凋亡调控基因，即C－myc基因，Bcl-2基因，P^53基因，ICE基因，Fas基因及FasL和Cdc2基因的来源，作用和相互间关系的研究近况，予以综述，做一简介。