绵羊FSTL3基因g.40674542C>T位点多态性与产羔数的关联分析

文章旨在探究绵羊FSTL3基因g.40674542C>T位点多态性与绵羊产羔数之间的关系,以期为绵羊高繁殖力分子育种提供新的遗传标记。利用全基因组重测序结合Sequenom MassARRAY~SNP技术对常年发情绵羊品种(小尾寒羊、策勒黑羊和湖羊)和季节性发情绵羊品种(滩羊、苏尼特羊和草原型藏羊)FSTL3基因g.40674542C>T位点多态性进行了检测,并与小尾寒羊产羔数进行了关联分析。结果表明:FSTL3基因g.40674542C>T位点存在CC、TC和TT三种基因型,基因型频率和等位基因频率在两种发情模式绵羊品种间差异均不显著(P>0.05);g.40674542C>T位点在6个绵羊品种中均表现为低度多态(PIC<0.25),经卡方适合性检验,该位点在小尾寒羊、湖羊和草原型藏羊中处于哈代温伯格平衡状态(P>0.05),在其余3个绵羊品种中处于哈代温伯格不平衡状态;g.40674542C>T位点不同基因型与小尾寒羊第一、第二以及第三胎产羔数均无显著关联(P>0.05)。说明FSTL3基因g.40674542C>T位点不适合用于小尾寒羊产羔数选育。     

弓形虫MIC3基因真核表达质粒的构建及在 IBRS-2细胞内的表达

采用PCR技术从弓形虫RH株的基因组DNA中扩增编码MIC3的基因,克隆入pMD18-T载体,转化至E.coliDH5α感受态细胞,经抗性平板筛选、小量抽提质粒进行酶切、PCR及DNA测序鉴定后,亚克隆入真核表达载体pcDNA3.1。然后筛选含有目的基因的重组质粒,并转染IBRS-2细胞,在G418压力下进行筛选,利用SDS-PAGE、Western blot和ELISA检测表达情况。结果显示,扩增的MIC3基因与GenBank上相应基因序列(AJ132530)的一致性达99.9%,构建的真核表达质粒pcMIC3能在转染的IBRS-2细胞中表达分子量约为39.2ku的MIC3,且表达的蛋白质具有良好的免疫活性,为进一步研究该质粒的动物免疫试验奠定了基础。     

基于"3S"技术的阿坝县退牧还草工程效益监测

应用“3S”技术(GIS、RS、GPS)为主要手段,辅以地面路线调查,通过建立遥感与地面数据的数学模型反演地面信息而进行了阿坝县退牧还草工程的遥感监测,并对工程实施情况、工程建设效益进行了评价。为进一步开展草原资源与生态动态监测和草原生态建设工程效益评价与监测等工作提供了技术模式。     

Antigen-nonspecific macrophage factors modulating the antibody response in vitro

Antibody response to an antigen involves the co-operation between three types of cells: macrophages, T cells and B cells. The cognate interactions between these cells play a fundamental role in the expression of a specific antibody response, but the last is modulated by antigen-nonspecific soluble factors produced either by macrophages or by T cells. Macrophages elaborate a spectrum of molecules modulating the function of lymphoid cells; among them are IL1 and prostaglandins of the E series, which are respectively enhancer and inhibitor of the antibody response in vitro. These molecules alter T cell and B cell activities through different mechanisms involving activation or inhibition of IL2 production, or alteration of cells surface antigens. However, the cellular events following the fixation of soluble factor on its receptors are not known.     

山羊碱性氨基酸转运载体基因SLC3A1cDNA序列克隆及表达

试验旨在克隆山羊碱性氨基酸转运载体基因SLC3A1 cDNA序列,探究其表达的组织特异性及其在小肠中的表达发育模式。参考GenBank已发表的绵羊、牛SLC3A1基因mRNA序列设计引物,克隆山羊SLC3A1基因的cDNA序列,采用实时荧光定量PCR的方法分析其在1日龄山羊11种组织及其在1日龄、6月龄、8月龄、10月龄、12月龄山羊十二指肠、空肠、回肠中的mRNA表达情况。结果表明,成功克隆得到了山羊SLC3A1基因cDNA序列,其与绵羊、牛、野猪、人、小鼠、大鼠的同源性分别为99%、97%、88%、86%、80%和79%。SLC3A1基因转录表达有明显的组织特异性,其在1日龄山羊肾脏、回肠、空肠、结肠中表达量依次降低(P<0.05),其他组织中表达量很低。同一月龄山羊,SLC3A1基因在不同肠段的表达量数值上回肠>空肠>十二指肠。SLC3A1基因在不同月龄山羊十二指肠表达量以1日龄山羊为最高(P<0.05),6~12月龄山羊相同肠段之间表达量无显著差异(P>0.05);空肠表达量随山羊年龄的增加均呈现逐步降低的趋势;回肠表达量在各个月龄山羊之间差异不显著(P>0.05)。结果说明,山羊碱性氨基酸转运载体基因SLC3A1的主要表达部位在肾脏和肠道,推测b^0,+碱性氨基酸转运系统转运氨基酸的主要部位为肾脏和肠道。SLC3A1基因在山羊小肠受肠段和发育阶段的影响,具有不同的表达发育模式。     

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.     

Effect of the β‐carotene oxygenase 2 genotype on the content of carotenoids,retinol and α‐tocopherol in the liver,fat and milk of rabbit does,reproduction parameters and kitten growth

Mutations in the β‐carotene oxygenase 2 (BCO2) gene can impair the function of the enzyme that breaks down carotenoids. As a result, gradual accumulation of unoxidized carotenoids in animal tissues gives them a yellow colour. The aim of the study was to determine the content of carotenoids, retinol and α‐tocopherol in the liver, fat and milk of rabbit does with three different genotypes determined by AAT‐deletion mutation at codon 248 of the BCO2 gene and to find out whether differences in the concentrations of the above compounds in the tissues and milk of the does affect reproduction parameters and the rearing rate of kittens. The experimental materials comprised 36 does, 12 of each genotype of the BCO2 gene, with their litters. Females with their litters were placed in individual cages, on deep litter. Between days 7 and 13 of lactation, samples of milk were collected from the does. The kittens stayed with their mothers until 35 days of age. After weaning, the does were sacrificed. Tissue samples of liver and perirenal fat were collected for chemical analyses. Additionally, based on samples taken from one female, RNA expression levels were determined from the mammary gland and liver, adipose tissue and skin. It was found that homozygous does with deletion at codon 248 of the BCO2 gene were characterized by considerably higher concentrations of xanthophylls and beta‐carotene in the liver, adipose tissue and milk than does with the remaining genotypes. However, the differences in the content of the above compounds in milk had no influence on litter weight or the number and rearing rate of kittens. Additionally, RNA expression of the BCO2 gene was found in the mammary tissue of lactating doe and its level was similar to those noted in the liver and adipose tissue.     

3H—萱草根素在小鼠体内吸收分布,排泄和代谢的初步观察

本文应用放射性同位素示综技术,初步观察了~3H-萱草根素在小鼠体内吸收分布、排泄和代谢的全过程。~3H-萱草根素口服后0.5小时,小鼠血液中放射性即达到高峰,并以低浓度维持96小时以上;其随血液循环分布于各组织器官,以胃肠组织含量最高,其次是肝脏和脾脏。萱草根素有蓄积性;一次给药和以同等剂量分次(每天一次,三天)投药后的相同时间,各组织中~3H-萱草根素的含量差异不显著。主要排泄途径是粪便,其次是尿液。经有机溶剂处理后的尿液分为水、脂二相,其放射性测量和薄板层析放射自显影结果均表明:~3H-萱草根素在体内经过代谢后结构发生了变化,从尿液中排出的不是萱草根素原形,而是其代谢产物。     

Expression and functional analysis of the Follistatin-like 3 (FSTL3) gene in the sheep ovary during the oestrous cycle

Follistatin-like 3 (FSTL3) is a regulator of cellular apoptosis and was previously identified via RNA-Seq to be associated with follicular development in mammalian ovaries. However, the mechanism underlying the FSTL3 regulation of oestrus in sheep remained poorly understood. In this study, the oestrogen (E2) and progesterone (P4) concentrations in blood were detected, and the expression level and functional analysis of FSTL3 in the ovary were studied during the different reproductive stage in Aohan fine wool sheep (seasonal breeding breed in China). The concentrations of E2 and P4 at the anestrus were significantly lower compared to dioestrus, proestrus and oestrus stages. Higher expression levels of FSTL3 were observed in the sheep ovary, hypothalamus, and thyroid. During different reproductive stages, higher expression levels were found during the stages of dioestrus and proestrus, while lower levels were found during the oestrus and anestrus stages. Functional analysis of FSTL3 was performed in primary granulosa cells (GCs) of sheep. The concentration of E2 increased significantly after RNAi interference of FSTL3, while the P4 level decreased. FSTL3 can decrease P4 levels, which might be involved in mediating oestrous cycle in sheep.