Expression and Bioinformatic Analysis of ADIPOQ Gene in Hanper Sheep

The aim of this study was to investigate the expression and biological characteristics of adiponectin (ADIPOQ) gene in longissimus dorsi muscle of Hanper sheep at different months of age.The ADIPOQ gene mRNA expression was detected in longissimus dorsi muscle of Hanper sheep at 3 different growth periods (1,7 and 13 months old) by Real-time quantitative PCR.The bioinformatics method was used to analyze the nucleotide and amino acid sequences of ADIPOQ gene,protein characteristics and network interaction protein.The results showed that ADIPOQ gene was expressed in longissimus dorsi muscle of Hanper sheep,which showed an increasing trend with the growth of the months age without significant difference (P>0.05).There was higher homology of ADIPOQ gene with Capra hircus (98.60%,99.00%) and Bos taurus (98.60%,87.00%) in nucleotide and amino acid sequences.The results of physical and chemical properties analysis showed that the molecular formula of ADIPOQ protein was C₁₁₇₂H₁₇₇₆N₃₁₄O₃₄₉S₅,the molecular mass was 26.00 ku,and the theoretical isoelectric point (pI) was 5.88,which suggested that the protein was acidic.The average value of ADIPOQ hydrophilic protein (GRAVY) was －0.403,which indicated that the protein was hydrophilic and didn’t belong to transmembrane protein.In addition,the signal peptide sequence was Met¹-Ala¹⁷,and the cleavage site was between Ala¹⁷ and Arg¹⁸.ADIPOQ existed collagen structure domain and C1Q conserved structure domain at position 45－101 and 101－237,respectively.The secondary structure mainly composed of random coil (66.95%).The construction of protein interaction network indicated that ADIPOQ might interact with ADIPOR1,ADIPOR2,CDH13 and LEP.The results provided a theoretical basis for further exploring the molecular biological functions of ADIPOQ gene in the process of muscle development in Hanper sheep.     

鲁梅克斯对草甸盐土的改土效应研究

在草甸盐土上种植鲁梅克斯Rumex patientia×R.tianschanicus cv.Rumex K-1,脱盐改土效果十分明显.种植鲁梅克斯3～4年,鲜草产量达100.51 t/hm2,pH值由8.43降为7.99,耕作层土壤总孔隙度增加5.29%～7.06%,土壤容重降低0.14～0.21 g/cm3,水稳性团粒结构增加23.14%～32.34%,自然含水量增加72.12～89.44 g/kg,土壤有机质、速效氮、磷、钾亦随之增加.     

加拿大披碱草-野大麦三倍体杂种加倍植株同工酶分析

分析了加拿大披碱草-野大麦三倍体属间杂种F₁加倍植株的同工酶酶谱特征。结果显示:同一生育阶段杂种自然加倍植株与加倍F₁代植株在EST、POD和SOD酶带的数目、位点及强弱方面具有一致性和遗传稳定性,而与杂种F₁代及亲本的酶带表型差异显著,从酶蛋白分子水平证明该杂种F₁染色体加倍是真实的;加倍植株抽穗期旗叶的EST、POD酶谱中分别呈现9和4条酶带,分蘖期幼叶的EST、POD、SOD酶谱内分别呈现8、4和4条酶带,有多态性位点的特征酶带可作为杂种加倍后代育性恢复鉴定的遗传标记的候选位点;对供试材料不同生育阶段做同工酶酶谱对比分析,比单一生育阶段更能反映其酶带表型的遗传差异性,提高同工酶电泳技术鉴定结果的准确性。     

鸡白介素18成熟蛋白的发酵工艺及其对新城疫疫苗的免疫增强作用

为探讨重组鸡白介素18（mChIL-18）毕赤酵母工程菌在5L发酵罐中大规模发酵的工艺及其对新城疫疫苗的免疫增强作用,复苏工程菌GS115/pPIC9K-mChIL-18于YPD培养基,在其D600值达到5.2左右时转入发酵罐中,采用分批补料方式对毕赤酵母工程菌进行高密度发酵。控制和优化各种发酵条件,经历84h结束发酵。将发酵液离心,用6×His镍柱对上清中的表达产物进行纯化,并将纯化后的mChIL-18用脂质体包被后和新城疫疫苗一起对鸡群免疫。结果显示,毕赤酵母工程菌在5L发酵罐采用甲醇诱导补料批式发酵,在pH5.5,溶氧值20%～30%,温度28℃,诱导72h,mChIL-18的表达产量为560mg/L,发酵上清经纯化后纯度可达70%以上;用0.2mL的mChIL-18脂质体能够显著增强机体的免疫力,其中HI抗体效价和淋巴细胞亚群CD4＋、CD8＋含量均显著高于对照组。这表明mChIL-18毕赤酵母工程菌在5L发酵罐高密度发酵成功,并能显著增强新城疫疫苗的免疫效果,为IL-18在生产实践得到更好的应用奠定了基础。     

Use of comparative proteomics to identify the effects of creatine pyruvate on lipid and protein metabolism in broiler chickens

Four hundred male chickens were selected to study the effects of pyruvate (Pyr), creatine pyruvate (CrPyr) and creatine (Cr) on the expression of hepatic mitochondrial and cytoplasm proteins associated with lipid and protein metabolism. Mitochondrial purification was accomplished using the two-step differential centrifugation and density gradient method, and the activities of organelle-specific marker enzymes were determined to assess the purity of the mitochondria. Proteins were extracted and fractionated by two-dimensional electrophoresis and the differential protein spots were assessed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. CrPyr reduced fatty acid accumulation by down-regulating adipose differentiation-related protein, inhibited ATP synthase expression, and reduced cholesteryl ester transfer protein (CETP) expression, thus reducing the levels of high density lipoprotein and triglycerol (TG) levels (thereby lowering fat and cholesterol deposition). CrPyr increased the expression of eukaryotic translation initiation factor (eIF) 2B, calreticulin (CRT) and eIF3a, thus promoting protein synthesis. CrPyr up-regulated the expression of fatty acid-binding proteins, CETP and apolipoprotein A-IV in cytoplasmic extracts, and these proteins accelerated the decomposition of fatty acids and TG, thus reducing fat deposition. In conclusion, CrPyr plays an important role in lipolysis and protein synthesis, and this effect was more pronounced than was the effect of Pyr and Cr.     

猪Ghrelin基因的克隆及原核表达

从猪下丘脑、胃等组织中提取总RNA，根据已发表的猪的Ghrelin mRNA序列设计合成引物，通过RT-PCR进行cDNA扩增，获得了282bp的片段。将该片段克隆于pMD-18T载体后进行序列分析，确认PCR产物为Ghrelin cDNA。从阳性克隆中提取质粒，经NheⅠ和XhoⅠ双酶切，回收282bp的目的片段，定向克隆到pET-28a表达载体中，提取质粒并再次转化到BL21（DE3）中，成功地筛选出阳性克隆。经IPTG诱导阳性菌，通过SDS-PAGE检测出猪Ghrelin基因的表达．     

月季种质资源花色基础研究

运用测色仪对227个月季种质资源花色表型进行分类,并运用紫外分光光度计测定总类黄酮,探讨二者之间的相关性。结果表明:在a~*和b~*二维象限图上,7类色系主要集中分布在Ⅰ、Ⅱ、Ⅳ象限,在第Ⅲ象限(蓝色)没有分布,花瓣正面颜色分布较为集中、部分橙红色和深红色品种的反面花色分布较为离散;7个色系的明度均为L~(*B)L~(*F),绝大多数色系的颜色参数a~(*F)/b~(*F)/C~(*F)a~(*B)/b~(*B)/C~(*B),并且玫红色、紫红色、橙红色和深红色的正反面花色a~*、b~*、C~*差异显著;各个色系TA之间差异显著(P0.05),深红色TA含量最高。而TF含量,除了深红色和紫红色差异显著外,其他色系之间差异不显著(P0.05);TA与L~*呈负相关关系(P0.01),与花瓣L~(*B)相关性更大;TA与h°负相关(P0.01)。TA与a~*/C~*呈正相关关系(P0.01),与正面颜色参数值相关性更强,而TF与L~*/h°也呈负相关关系(P0.05)。说明TA和TF共同影响花色的呈现,并且花青苷含量的高低是影响花色深浅的主要因素。     

全膜覆土穴播粒数对紫花苜蓿生产性能的影响

采取随机区组排列的方式,设置3种不同穴播粒数,对陇中苜蓿和岩石进行生长高度、产草量、茎叶比、粗蛋白含量等指标测定,结果表明:穴播粒数为14时,陇中苜蓿和岩石的生长高度表现较为突出,鲜草产量分别为43 404.5kg/hm~2和47 436.9kg/hm~2,显著高于其它两个处理;在品质方面也表现较好,粗蛋白含量分别为15.33%和14.74%。综合评价全膜覆土穴播苜蓿14为最佳穴播粒数。     

草地生态系统安全及其评价研究

探讨了草地生态安全及其评价的概念与内涵,认为草地生态系统安全判断与度量准则应该是草地生态系统健康、草地生态系统服务功能及生态风险与管理的科学统一。同时尝试提出了草地生态系统安全评价的"健康-服务功能-风险与管理(health-service-risk and management,HSRM)"模型,构建了HSRM评价模型的指标体系,提出草地生态安全评价中草地生态安全度计算公式和安全状态分级标准。