牛体外受精时老化卵对受精率与染色体异常发生的影响

通过延长体外培养时间得到牛老化卵，对其体外受精率与染色体的异常发生进行了研究，体外受精48h后的受精率，46h培养组与22h和34h培养组之间差异显著（P＜0．05），随着老化时间的延长，受精率及胚胎发育速度明显降低。染色体分析结果表明，46h组的受精卵的染色体异常发生率为61．6％，22h组44．6％，2组之间差异显著（P＜0．05），单倍体（n=31)的发生率也随着老化时间的延长而增加，这些单倍体的性染色体的构成表明，含有X－性染色体的胚的出现率显著高于含有Y－染色体的胚的出现率（P＜0．05）。单倍体的发生是由卵的老化而引起单一配子的孤雌发生，体外培养时间的延长对正常的2倍体的性别无明显影响。     

沼泽型水牛ALK3基因克隆分析与表达模式研究

试验旨在对沼泽型水牛ALK3基因进行克隆、生物信息学分析,并对其在水牛组织中的表达规律进行系统研究.根据GenBank中已公布的牛ALK3基因序列设计特异性引物,应用RT-PCR方法扩增、克隆获得目的基因片段;应用生物信息学方法分析和预测了水牛ALK3的遗传进化及蛋白质的理化性质、二级和三级结构;并应用QRT-PCR技术对ALK3基因在水牛组织中的表达进行了差异分析.结果表明,水牛ALK3基因编码区全长1 599 bp,共编码532个氨基酸.多重序列比较分析显示,水牛ALK3核苷酸序列与牛、绵羊、猪、马、人和小鼠相应序列的同源性分别为98%、96%、95%、93%、94%和91%.系统进化树分析显示,ALK3基因在不同物种以及进化的过程中具有高度保守性.对ALK3蛋白质的分析表明该蛋白呈弱碱性,有信号肽,细胞亚定位于胞膜上,存在丝氨酸/苏氨酸激酶和GS等结构.定量分析结果显示,ALK3在水牛生殖脊、心脏、肝脏、颗粒细胞、肺脏、卵丘细胞、肾脏、下丘脑、垂体、大脑等15种组织或细胞中有不同程度的表达,其中卵巢中表达量最高,垂体、肺脏和睾丸次之,卵丘细胞表达量最低.本研究成功克隆了沼泽型水牛ALK3基因,并研究了其在不同水牛组织细胞中的表达规律,为阐明其在水牛繁殖过程中的功能及转基因载体构建中的应用研究奠定了理论基础.     

杀菌型乳酸菌饮料的模糊数学综合感官评判方法

通过对市场上流行的5种杀菌型乳酸菌饮料的焦糖味、果味、发酵味、甜味、酸味、涩感的感官评定,采用组合赋权法得出杀菌型乳酸菌饮料焦糖味、果味、发酵味、甜味、酸味、涩感的质量权重.对5种杀菌型褐色乳酸菌饮料各指标以权重,然后结合模糊数学综合评判方法,得出模糊关系曲线.结果表明:5种杀菌型乳酸菌饮料的整体感官排序为:样品5＞样品2＞样品4=样品l＞样品3,与一般排序法所得结果样品2＞样品4=样品1＞样品5＞样品3相比较稍有不同.因此,模糊数学综合评判方法比常用的排序法更科学、合理、客观.     

西藏兔病毒性出血症的研究--病原鉴定及疫苗免疫效力试验

１９９４年６￣７月,西藏林芝地区家兔大批急性死亡同,经过流行病学调查,临诊断症状观察,病理剖检等结果与兔病毒性出血症特征相符。进一步作病毒学检查表明,病变组织超薄发切片经电子显微镜观察发现有大量直径约３０ｎｍ的球状裸听病毒粒子,自然病死兔肝组织匀交涉 清能凝集入）型红细胞,而且血凝兔出血症病毒阳性抗血清所抑制,致病性试验结果表明,该病毒对兔有很强的致病性;用西藏自然病死兔肝组织匀浆试制的矩形疫苗兔     

Vero E6细胞酵母双杂交cDNA文库的构建及鉴定

To seek out proteins which interact with structural proteins of porcine epidemic diarrhea virus （PEDV） in Vero E6 cells and study the mechanisms of viral infection.We created a Vero E6 cDNA yeast hybrid library. The total RNA of Vero E6 was extracted using Trizol method, and double strands cDNA was synthesized by SMART technique. We created a Vero E6 cDNA library by using homologous recombination in yeast. The results showed that the titer of cDNA library was 9.7×10⁸ CFU with the average of inserted fragments about 1.6 kb, and the recombination rate was 87%. These data indicated that the yeast two-hybrid cDNA library of Vero E6 was successfully constructed and might be useful for screening the host proteins interacting with structure protein of PEDV.     

Cloning and Sequence Analysis of Major Histocompatibility Complex Ⅰ Gene from Goose F1 of Fast-growth Lines

According to the multiple alignments identified major histocompatibility complex Ⅰ (MHC Ⅰ) gene conserved sequence registered in GenBank from the family ducks (Anatidae) anser waterfowl, a pairs of specific primers for the fragments of MHCⅠgene of goose F1 from fast-growth lines were designed and synthesized by Primer Premier 5.0. Using the genome DNA of goose F1 from fast-growth lines, the target gene fragment was obtained by PCR. To conduct sequencing of the fragments of MHCⅠgene of goose F1 from fast-growth lines and make sequence alignment and analysis of protein structure and function by bioinformatics, and research the characteristics of MHCⅠgene of goose F1 and the physicochemical properties of the protein. Bioinformatics was analyzed the nucleic acid data, deduced amino acid sequence and phylogenetic trees. The result of sequence analysis showed that the fragments of MHCⅠgene of goose F1 from fast-growth lines was 1036 bp in length, which coded 96 amino acids polyprotein. The homology were 93% and 83% with MHC Ⅰ gene and coding sequence of Wulong goose in NCBI respectively. There were 72 different bases sequence and 16 amino acids change. There also was higher homology with other poultry, and existed genetic relationship of Siji goose > chickens > ducks.The homology segment sequences corresponding to the fragments of MHCⅠ gene of goose F1 coded 96 amino acids protein, which molecular weight, PI, positively or negatively charged amino acid, estimated half-life, instability index, aliphatic index and average hydrophobicity were 11.342 ku, 5.32, 14, 17, 2.8 h, 34.92, 42.81, -1.066, respectively, and appeared 9 B cell epitopes, but contained no signal peptide. These results indicated that the protein for hydrophilic non-secreted proteins, had the high immunogenicity. In addition, The protein structure study indicated that alpha-helix, beta-sheet, beta-turn and random coil were 31.25%,16.67%, 14.58% and 37.50%, respectively. There existed amino terminal domain and carboxyl terminal domain in the tertiary structure. Therefore, MHC gene had significant difference between species and populations of individuals by the pathogen pressure in environment, and there were the interaction between polymorphism of MHC molecules and the diversity of antigenic peptide. MHC determined the differences of individual susceptibility to disease, and could be treated as a candidate gene for disease resistance.     

中度盐碱化草地混播牧草替代玉米青贮对绵羊生产性能的影响

为研究中度盐碱化草地混播牧草对绵羊采食量、日增重和屠宰性能的影响,试验选用24只3月龄、体重(23.1±0.62) kg的德国肉用美利奴杂交一代绵羊,随机分为4组,对照组饲喂精料补充料十玉米(Zea mays)青贮,处理Ⅰ,Ⅱ和Ⅲ组在对照日粮基础上分别以中度盐碱化草地混播牧草(披碱草(Elymus dahuricus)、碱茅(Puccinellia distans)和沙打旺(Astragalus adsurgens))替代1/3,2/3和3/3玉米青贮,每只羊精料补充料日喂600 g,粗饲料自由采食.结果表明:处理Ⅰ,Ⅱ和Ⅲ组的有机物、干物质、酸性洗涤纤维、中性洗涤纤维采食量均显著高于对照组(P＜0.05);处理Ⅰ和Ⅱ组的日增重虽略高于对照组,但差异不显著;处理Ⅱ和Ⅲ组的饲料转化率显著高于对照组;各处理组绵羊屠宰体重、胴体重、屠宰率、净肉率、胴体出肉率、肉骨比、眼肌面积均较对照显著提高(P＜0.05).因此,中度盐碱化草地混播牧草替代玉米青贮可以增加绵羊的采食量,提高绵羊的屠宰性能,并以处理Ⅰ和Ⅱ组的效果较理想.     

24％溴虫腈·辛硫磷乳油对家蚕残毒试验

桑园治虫用药一直移用农田农药,安全性差,家蚕中毒事故频发;近年虽有一些桑园专用农药推出,但由于数量少,使用频率高,促成了害虫耐药性迅速提高,造成多种药物防效明显下降;开发桑园专用新农药,对桑园害虫控制和抗性治理都有积极意义。     

裂区设计下分析影响欧洲菊苣产量的因素

在裂区设计下,分析影响欧洲菊苣（Cichorium intybus cv.Europ）产量的7个因素,结果表明：播种季节对株高、产量有显著性影响;播种方式对株高、茎粗、密度、盖度、产量5个指标均有显著影响。进一步通过因子分析得出欧洲菊苣产量最佳组合为撒播、密度在50～59株·m^-2、盖度为94%～100%。     

桦木酸抑制地塞米松诱导脾淋巴细胞凋亡的作用

研究桦木酸对脾淋巴细胞凋亡的影响。以地塞米松建立小鼠体外脾淋巴细胞凋亡模型,利用倒置显微镜、荧光显微镜和电子透射显微镜观察细胞形态和亚细胞结构,琼脂糖凝胶电泳观察DNA的裂解,流式细胞术测定细胞凋亡率,检测华木酸对Dex介导的小鼠脾淋巴细胞凋亡的影响。结果表明,2.5μmol Dex可使脾淋巴细胞凋亡明显增加,染色质发生边聚、浓缩、凋亡小体等典型的凋亡特征,桦木酸可明显降低地塞米松引起的脾淋巴细胞凋亡。提示桦木酸对地塞米松诱导的淋巴细胞凋亡具有预防性保护作用。