家鸡Leptin成熟肽cDNA的克隆、重组蛋白表达及纯化

从 18周龄鸡卵巢组织中抽提总 RNA,使用六聚体随机引物反转录后 ,用鸡 L eptin(瘦素 )特异性引物扩增出鸡L eptin编码区第 5 2～ 4 6 0 bp的长度为 4 0 9bp的 c DNA片段。根据鸡 L eptin的 3′端第 4 6 0～ 4 92 bp序列设计了 3条部分相互重叠并且顺序串联延伸的下游反义引物 ,并在最后 1条引物 3′端连接上 Eco R 切点 ;在以上用于反转录扩增的 5′端引物的 5′端连接一 Bam H 切点。用该 5′端引物分别与 3个 3′端引物配对 ,利用反转录扩增出的 4 0 9bp的L eptin c DNA片段作为第 1模板进行扩增 ,扩增产物再作为模板与下一引物对再次扩增。经 3次扩增得到编码鸡 L ep-tin成熟肽的全长 4 5 1bp的 c DNA序列。将该 4 5 1bp的 L eptin c DNA序列经 Bam H 和 Eco R 双酶切后 ,克隆入表达质粒 p RSET A的 Bam H 和 Eco R 两酶切位点之间 ,构建成表达质粒 p L ep- SCAU。转化有重组表达质粒 p L ep-SCAU的大肠杆菌 BL 2 1(DE3)在 L B培养基中培养后 ,经 IPTG诱导表达出相对分子质量为 2 0 10 0的鸡 L eptin融合蛋白和少量 4 0 2 0 0的 L eptin融合蛋白。L eptin融合蛋白的表达在 IPTG浓度为 0 .0 5 mmol/ L 时达到最高 ,占总菌体蛋白的 32 .6 %。用 Ni- NTA凝胶从 7L 发酵培养菌裂解液中纯化出 180 m g左右     

百例牛剖腹产手术的经验及技术探讨

难产是牛常见的产科病之一，奶牛发病明显高于黄牛。难产发生以后，经过各种办法难以从阴道产出犊牛，就应果断的，不失时机的施行剖腹产手术，从而有效的救治牛只。如能慎重果断的选择病例，早期进行手术，加上熟练的技术，成功率是很高的。     

华北农牧交错带退化草地土壤种子库动态变化

对华北农牧交错带不同退化演替阶段土壤种子库的数量和物种组成及与地上植被的关系进行了初步的研究,结果表明:农牧交错带草地在刈割和放牧等干扰下,发生逆行演替,土壤种子库中植物种类及种子数量呈递减的趋势,轻度退化类型土壤种子库物种组成最高,有23种植物,种子数量最多,有4 303粒,禾本科所占比例较大,为32.3%;而极度退化草地物种组成最低,有14种植物,种子数量仅有1 156粒,禾本科只占8.4%.草地逆行演替过程中植被与土壤种子库在物种组成上存在一定差别,中度退化阶段差别最大,有18种植物未在种子库中出现,极度退化阶段差别最小,有2种未在种子库中出现.     

Rapid detection of contagious ecthyma by loop-mediated isothermal amplification and epidemiology in Jilin Province China

The aim of this experiment was to develop a loop-mediated isothermal amplification (LAMP) assay and to research the recent epidemiology of contagious ecthyma in Jilin Province, China, using the assay. A LAMP assay targeting a highly conserved region of the F1L gene was developed to detect contagious ecthyma virus (CEV). Three hundred and sixty-five cases from 64 flocks in 9 different areas of Jilin Province, China, from 2011 to 2014 were tested using the LAMP assay. The results showed that the sensitivity of the LAMP assay was 100 copies of the standard plasmid, which is 100-fold higher than the sensitivity of PCR. No cross-reactivity was observed with capripoxvirus, fowlpox virus, foot-and-mouth disease virus serotype O, foot-and-mouth disease virus serotype Asia I and bluetongue virus. The average positive rate was 19.73% (72/365), and the positive rate was highest in lambs aged 1–6 months. Our results demonstrated that CEV infection was very widespread in the flocks of Jilin Province and that the LAMP assay allows for easy, rapid, accurate and sensitive detection of CEV infection.     

蜂王浆对尾静脉注射4T1细胞小鼠抗氧化能力的影响

探讨蜂王浆对尾静脉注射4T1细胞小鼠的抗氧化和免疫力的影响.将4T1细胞或其与蜂王浆的混合物通过尾静脉注射入小鼠体内,ELISA法检测蜂王浆对小鼠血清、肝脏和肾脏中与抗氧化能力相关的指标及血清中和免疫力相关细胞因子水平的影响.结果表明,和模型对照组相比,蜂王浆(50、100 mg·mL~(-1))可以显著提高小鼠血清内T-AOC、IFN-α(P0.05)水平,降低IL-4的水平(P0.05);蜂王浆剂量为50 mg·m L~(-1)时,IFN-γ的含量显著增加(P0.05).蜂王浆(100 mg·mL~(-1))显著提高肾脏中T-AOC的水平(P0.05).可见,蜂王浆可以改善尾静脉注射4T1细胞小鼠的抗氧化能力和免疫力.     

地裂缝构造背景与坳陷伸展构造环境的关系

首次从坳陷伸展构造环境的角度出发,分析和总结了前人对冀中坳陷伸展构造环境的研究和系统的建立,探讨了6处河北平原典型构造成因地裂缝发育带区域地质背景和伸展构造环境,认为伸展构造环境为地裂缝发育提供了内部空间、外源动力等条件,并控制了地裂缝发育区域与主要特征。在此环境下,地裂缝的产生是伸展构造环境下第四系土体演化过程中的地质现象之一,地裂缝的差异性主要是由基底起伏、伸展断层(变换断层)、第四系覆盖土体的物理性质、人类工程活动等内外因素造成的。     

Identification and Molecular Characteristics Analysis of Porcine Epidemic Diarrhea Virus Variants

To ascertain a diarrhea case in a pig farm in Shandong province,the pathological changes of dead piglets were observed and nested RT-PCR test was carried out on 7 diarrhea samples for porcine epidemic diarrhea virus (PEDV).Pathological examination revealed that intestine was detected as enlargement,hyperemia and edema.After histological examination,the typical microscopic lesions of intestine were disappearance of epithelial cells,villus shrinkage and shortening.The result of nested RT-PCR showed that all of the 7 samples could amplify a specific target band of PEDV.Molecular characteristics of two field strains showed that they had an amino acid homology of 92.3% to 92.4% with vaccine CV777 and 96.6% to 98.6% with other previous field strains which sequences were downloaded from GenBank.Phylogenetic tree analysis further revealed that all of PEDV strains could be mainly divided into two clusters of G1 and G2. G2 consisted of the field strains of our study and other field strains from USA,China and so on,which had the same sequence characteristics of two insertions and one deletion,while G1 consisted of all vaccines of CV777 and several older field strains from China and Korea.These results indicated that our field strains were the dominant strains in recent epidemic diarrhea occurrence.Moreover,its molecular characteristics might be a characterization of differentiating the field and vaccine strains.     

食品中氨基酸的作用及测定方法

氨基酸是生物功能大分子蛋白质的基本组成单位,被广泛应用于食品、医药、保健品等行业.其研究和测定方法一直是人们关注和研究的热点,文章较全面地阐述了食品中氨基酸的作用及相关测定方法.     

内蒙古牧区现代化建设中畜牧业高质量发展对策的研究

在对内蒙古部分牧业旗（县）实地调研的基础上,总结分析以33个牧业旗（县）为代表的内蒙古牧区畜牧业发展取得的进步和存在的主要问题,系统地提出了注重产业化经营、注重规划专业化生产布局、注重草原畜牧业生产方式转变、注重良种繁育体系建设、注重调整畜群结构及繁育方式、注重农牧区饲草资源互补利用体系建设的“六注重”牧区畜牧业高质量发展建议,以期为推进内蒙古牧区现代化建设提供参考。     

砂田甜瓜套作向日葵系统的养分利用研究

2010—2013年在庄浪县进行的砂田甜瓜套作向日葵系统的养分利用研究的结果表明,无论在甜瓜的初花期、座果期、膨大期套作食葵或油葵,套作系统比单作明显提高了作物的吸氮量、吸磷量和吸钾量。氮吸收量比单作增加50%~59%,磷吸收量比单作增加12%~41%,钾吸收量比单作增加33%~68%。