Cloning and Tissue Expression Analysis of PDK4 and FGF10 Genes in Guizhou Congjiang Xiang Pig

The objective of this study was to clone PDK4 and FGF10 genes, and investigate the expression level of PDK4 and FGF10 genes mRNA in different tissues of Large White pig and Congjiang Xiang pig. The PDK4 and FGF10 genes were cloned by RT-PCR and analyzed by bioinformatics, the relative expression of PDK4 and FGF10 genes were detected by Real-time PCR. The results showed that the coding region of PDK4 gene was 1 224 bp, encoding 407 amino acids; The coding region of FGF10 gene was 636 bp and encoded 211 amino acids. The homologies of nucleotide sequences of PDK4 gene with sheep, horse and human were 93%, 92% and 91%,respectively. The homologies of nucleotide sequences of FGF10 gene with sheep, cattle, human and mouse were 94%,93%, 93% and 90%, respectively. The phylogenetic tree of PDK4 gene showed that the genetic relationship of Congjiang Xiang pig, cattle and sheep were very close, the phylogenetic tree of FGF10 gene indicated that the genetic relationship of Congjiang Xiang pig, cattle, sheep, human and macaque were very close, but the genetic relationship of Congjiang Xiang pig, rat and chicken were far away. Real-time PCR results showed that, in different tissues of Congjiang Xiang pig,PDK4 gene expression in kidney tissue was higher than other tissues, with a higher expression in stomach and adipose as well,FGF10 gene expression in stomach tissue was higher than other tissues, with a higher expression in kidney and adipose as well, but both of PDK4 and FGF10 genes expression were the lowest in longissimus dorsi. In different tissues of Large White pig, both of PDK4 and FGF10 genes were expressed the highest in adipose than other tissues, PDK4 gene expression in longissimus dorsi was the lowest, while the FGF10 gene expression the lowest in heart. This study successfully cloned the PDK4 and FGF10 genes of Large White pig and Congjiang Xiang pig,and detected the relative expression of PDK4 and FGF10 genes in different tissues of Large White pig and Congjiang Xiang pig, and also provided scientific basis for further study on regulation of PDK4 and FGF10 genes on lipid metabolism and deposition.     

降解赤霉病菌毒素Tri101基因的原核表达及抗体制备

采用RT-PCR方法克隆了编码禾谷镰刀菌单端孢酶烯3-O-乙酰转移酶Tri101基因的cDNA序列,并连接到原核表达载体pGEX-4T2上,将获得的重组载体pGEX-4T2/Tri101转化大肠杆菌BL21后用IPTG进行诱导表达。SDS-PAGE和Western blot分析表明,经IPTG诱导后,Tri101基因在大肠杆菌BL21中获得了高效表达,融合蛋白GST-Tri101分子量为75.45 kDa。将该融合蛋白切胶纯化后免疫家兔,制备兔抗GST-Tri101多克隆抗体。经ELISA法测定抗体效价大于1∶256 000。Western blot分析表明制备的抗体与原核细胞体外表达的Tri101蛋白可以特异性结合,表明该抗体的特异性良好。应用该抗体验证了感赤霉病小麦中Tri101基因的表达。兔抗GST-Tri101抗体的成功制备,为进一步研究Tri101的生物学功能、细胞定位以及在其它植物中的表达等奠定了基础。     

苹果砧木M26试管苗不定根发育过程中蛋白质的变化

为进一步了解木本植物不定根发生、发育的机理,对苹果砧木M26试管苗不定根的发生、发育过程进行解剖观察,利用十二烷基磺酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)对M26试管苗在接入生根培养基前后,不同时期表达的蛋白进行了研究。结果发现:在不定根原基形成和发育的阶段性过程中,茎基部蛋白质的种类和数量出现明显差异。生根培养后,在继代苗茎基部表达的5条蛋白条带减弱或消失,其分子量分别为26.8 kDa、37kDa、40.3 kDa、43 kDa和66 kDa;明显表达了4条新的蛋白条带,其分子量分别为26 kDa、27.7 kDa、32.5 kDa、和45 kDa,同时有4条蛋白条带(38.5 kDa、42 kDa、53.2 kDa和55.5 kDa)其表达丰度提高。多种蛋白条带的消失和出现,可作为M26试管苗不定根产生的生化标志。在不定根原基逐步形成发育的同时,有3条蛋白条带的表达丰度也逐步提高,可作为不定根形成过程表达的特异标记蛋白带。     

Physiological but not morphological adjustments along latitudinal gradients in a human commensal species,the Eurasian tree sparrow

Human commensal species take advantage of anthropogenic conditions that are less likely to be challenged by the selective pressures of natural environments. Their morphological and physiological phenotypes can therefore dissociate from habitat characteristics. Understanding how these species adjust their morphological and physiological traits across latitudinal gradients is fundamental to uncovering the eco-physiological strategies underlying coping mechanisms. Here, we studied morphological traits in breeding Eurasian tree sparrows (ETSs; Passer montanus) among low-latitude (Yunnan and Hunan) and middle-latitude (Hebei) localities in China. We then compared body mass; lengths of bill, tarsometatarsus, wing, total body, and tail feather; and baseline and capture stress-induced levels of plasma corticosterone (CORT) and the metabolites including glucose (Glu), total triglyceride (TG), free fatty acid (FFA), total protein, and uric acid (UA). None of the measured morphological parameters varied with latitude except in the Hunan population, which demonstrated longer bills than those in other populations. Stress-induced CORT levels significantly exceeded baseline levels and decreased with increasing latitude, but total integrated CORT levels did not vary with latitude. Capture stress-induced significantly increased Glu levels and decreased TG levels, independent of site. However, the Hunan population had significantly higher baseline CORT, baseline and stress-induced FFA levels, but lower UA levels, which differed from other populations. Our results suggest that rather than morphological adjustments, physiological adjustments are mainly involved in coping mechanisms for middle-latitude adaptation in ETSs. It is worth investigating whether other avian species also exhibit such dissociation from external morphological designs while depending on physiological adjustments.     

奶牛粪样中致病性大肠埃希菌的分离与鉴定

大肠埃希菌血清型众多,一些具有特殊血清型的大肠埃希菌对人和动物有致病性。本试验从包头市侯家营奶牛场77头外表健康成年泌乳荷斯坦奶牛采集新鲜粪样,进行细菌分离培养,共分离到疑似大肠埃希菌69株,经过形态观察、生化鉴定、动物致病性试验和血清型鉴定,其中12株为致病性大肠埃希菌,均为强毒株,占分离菌数的17.4%,另57株为非致病性大肠埃希菌,占分离菌数的82.6%。同时对12株致病性大肠埃希菌进行了"O"血清型鉴定,分属7种血清型O78、O6、O9、O98、O1、O8、O149,分别占定型菌株的25.0%(3/12)、8.3%(1/12)、16.7%(2/12)、8.3%(1/12)、8.3%(1/12)、8.3%(1/12)、25.0%(3/12)。     

驻马店小麦条锈病发生气象指数预报方法探讨

小麦条锈病是河南省驻马店市小麦生产中的主要病害之一,历史上曾多次造成小麦严重减产。在分析小麦条锈病发病及流行特征的基础上,应用统计方法建立了条锈病病发气象指数预报气候模型和指数预测方程,并连续2a准确作出病发气象指数预报,为植保部门正确作出病害流行趋势预测预报提供了科学依据。     

皖北石灰岩山地不同植被恢复模式对植物群落物种组成及多样性的影响

通过研究皖北石灰岩山地生态系统的植被群落,分析不同植被恢复群落的组成、结构、多样性和生活型组成,进而阐明石灰岩山地生态系统植被恢复模式对群落结构特征以及植物物种多样性的影响。结果表明:研究区有54种植物,隶属26科54属。侧柏(Platycladus orientalis)+构树(Broussonetia papyrifera)人工混交林有植物种类16科26种,分别占总数的61.5%和48.1%。酸枣(Ziziphus jujuba var. spinosa)+牡荆(Vitex negundo var. cannabifolia)灌丛和荩草(Arthraxon hispidus)+牡荆草灌丛植物种类分别为32和33种,分别占调查总物种数的59.3%和61.1%。不同植被恢复群落生活型也有差异。侧柏+构树人工混交林群落高位芽植物占优势,而酸枣+牡荆灌丛和荩草+牡荆草灌丛则以地面芽植物所占比例较大。分析表明,多样性指数、均匀度指数和优势度指数均表现为侧柏+构树混交林>酸枣+牡荆灌丛>荩草+牡荆草灌丛,但物种丰富度表现为荩草+牡荆草灌丛>酸枣+牡荆灌丛>侧柏+构树混交...     

2013年沈阳地区汛期水情趋势分析

针对沈阳地区旱涝发生的基本规律,根据多年来开展水文中长期预报所应用的理论和方法,总结水情趋势预测工作的经验,对2013年水情趋势进行分析预测。     

高通量测序分析中国沙棘根瘤与根际土壤细菌多样性

以中国沙棘根瘤和其根际土壤为材料,采用高通量测序技术,分析沙棘根瘤内生细菌与根际土壤细菌的多样性差异,探究沙棘根瘤中引起根部瘤状突起的以弗兰克氏菌(Frankia)为主的沙棘根瘤内生细菌的来源。结果表明:沙棘根瘤中检测到的内生细菌归属于10门23纲45目76科和93属,其中,优势菌门为Actinobacteria和Proteobacteria,优势菌属为Frankia。在沙棘根际土壤中检测到的细菌归属于39门84纲156目280科和409属,其中,优势菌门为Proteobacteria、Actinobacteria和Acidobacteria,优势菌属为Gaiella、Bacillus、Solirubrobacter、Arthrobacter和Stenotrophomonas。可见,中国沙棘根际土壤细菌多样性要高于根瘤内生细菌多样性,沙棘根瘤内生细菌与根际土壤细菌的群落结构既有相似性又存在差异,其中,在根瘤中占优势的Frankia在根际土壤中虽也存在但相对丰度较小,仅为0.02%。     

AsiaⅠ型口蹄疫病毒VP1基因在昆虫细胞/杆状病毒中的表达

利用杆状病毒表达系统对Asia Ⅰ型口蹄疫病毒(foot-and-mouth disease virus,FMDV)VPl基因在Sf9昆虫细胞中进行表达,为研究Asia Ⅰ型FMDV VP1蛋白功能及建立Asia Ⅰ型FMDV血清学诊断方法奠定基础.采用PCR方法从pGEM-T-Easy-Asia Ⅰ型VP1质粒中扩增VP1基因,将其插入杆状病毒转座载体pFastBacHTA,构建的重组质粒pFast-BaeHTA-VPl再转入DH10Bae感受态细胞,经三重抗性与蓝白斑筛选,获得杆状病毒重组质粒Baemid-VPl,然后转染Sf9昆虫细胞.PCR鉴定证实VP1基因正确地插入到Bacmid中,成功构建了杆状病毒重组质粒Baemid-VP1,SDS-PAGE和West-ern-blotting检测结果表明,VP1基因在Sf9昆虫细胞中表达出约26.5 ku的VP1蛋白.将可溶性表达的融合蛋白用Ni-NTA亲和层析方法进行纯化,通过ELISA分析,能特异性地检测出Asia Ⅰ型口蹄疫病毒阳性血清.Asia Ⅰ型FMDV VP1基因在杆状病毒表达系统中的成功表达为Asia Ⅰ型FMDV VP1蛋白的抗原性及血清学抗体水平检测研究奠定了基础.