真核表达载体构建表达ＰＲＶ闽Ａ株gE基因表位抗原编码区片段的重组质粒

根据伪狂犬病病毒闽Ａ株gE基因表位抗原编码区的序列与身份种真核表达载体pPICZaA、pAcGP67A序列与特性分别设计了两对ＰＣＲ引物。通过ＰＣＲ方法扩增到了两端具有不同酶切位点的gE基因表位抗原编码片段。将这２个片段分别克隆到pPICZaA与pAcGP67A载体，转化大肠杆菌ＴＯＰ１０菌档及ＸＬ１－Ｂlue菌株，获得了含伪狂犬病病毒闽Ａ株gE基因表位抗原编码区的重组质粒pICZaA-FS与pAcGP67A-FS。序测定结果显示两个重组质粒中插入片段的大小与方向均正确。     

家畜繁殖生物技术研究现状及趋势

牛胚胎移植技术已趋于成熟。我国新疆牧科院用一步细管法移植牛冻胚受胎率达41%;牛和羊鲜胚四分胚移植也产下1头犊牛和6只羔羊。家畜体外受精,因卵子体外成熟和受精卵体外培养尚不过关,目前仍停留在实验室阶段。胚胎性别鉴定,1990年Koopman发现单拷贝基因,该基因是Y染色体的性决定区,命名为SRY,可利用PCR技术制成雄性特异DNA探针盒,进行马、牛、羊、猪早期胚胎性别鉴定。北京农学院等单位用PCR扩增牛SRY序列进行奶牛胚胎性别鉴定准确率达100%。英、日、法等国已获得牛胚胎细胞核移植后代;我国也获得1只核移植兔。北农大和新疆牧科院合作以绵羊精子为载体导入牛生长激素基因rMTbGH DNA成功,外源基因整合率为3%。     

生殖激素产品的应用和发展趋势

生殖激素是调控动物体内复杂生殖过程并影响繁殖性能的一系列的重要激素.近年来,利用生殖激素的批次化生产繁殖技术被广泛使用,提高了母猪的繁殖效率,促进了养猪业向集约化和规模化方向的发展.目前在批次化生产过程中主要采用孕马血清促性腺激素(PMSG)和人绒毛膜促性腺激素(HCG)以及化学类激素进行繁殖调控.但是这些产品的应用仍...     

Dvl1及其突变体腺病毒载体的构建与在MSCs中表达鉴定

Dvl1及其结构缺失突变体ΔDIX（dsh and axin）和ΔDEP（dsh,Egl-10 and pleckstrin）腺病毒载体的构建并验证其在MSCs（mesenchymal stem cells）中的表达情况。将目的基因定向克隆至pAdTrack-CMV穿梭载体上,并经PmeⅠ线性化后在BJ5183细菌中与pAdEasy-1骨架质粒同源重组,获得重组腺病毒载体,在QBI-293A细胞中包装及扩增,实时荧光定量PCR和Western bolt验证Dvl1在MSCs中的表达情况。经PCR、PacⅠ单酶切鉴定及测序分析,成功构建Ad-Dvl1、Ad-ΔDIX和Ad-ΔDEP腺病毒载体,目的基因序列与GenBank报道一致,并选出150 MOI为最适感染MSCs的感染复数,成功构建了Ad-Dvl1、Ad-ΔDIX和Ad-ΔDEP腺病毒载体,并获得高滴度的病毒子,qPCR和Western blot证实Dvl1在MSCs中高表达并增加了β-catenin在细胞中的累积,为进一步研究Dvl1在MSCs迁移过程的作用奠定了基础。     

重组白细胞介素-2高效纯化方法建立

依次通过离子交换法、凝胶过滤法纯化重组白细胞介素-2(IL-2)粗品,利用SDS-PAGE电泳和hIL-2 Elisa kit检测纯化后重组IL-2的纯度和浓度。结果表明,纯化后的重组IL-2纯度和浓度高。本纯化方法简单、稳定、纯化效果好,可为重组IL-2的生产工艺提供技术支持和依据。     

Genetic analysis and QTL mapping of growth period traits and plant height traits in soybean recombinant inbred lines from Dongnong 47 × PI 317334-B

High and stable yield is the main goal of soybean genetic improvement. In this study, association analysis was used to detect the quantitative trait loci (QTL) for the plant height, and soybean growth period using 182 SSR markers in the RIL population of 136 F_4:8 lines, which developed from a cross between photoperiod-insensitive cultivar ‘Dongnong 47’ and photoperiod-sensitive variety PI317334–B. The results showed that 33 QTLs related to soybean growth period and plant height traits were detected by compound interval mapping, and were located on 12 linkage groups including N, C1, C2, J, D1a, B2, E, G, A2, O, L, I, with the contribution rate of 7.85–33.84%. These QTL loci and linkage markers related to soybean photoperiod sensitivity, would be helpful to identify key genes that control soybean photoperiod sensitivity, and provide an important basis for the breeding of new photoperiod-insensitive soybean varieties based on molecular design breeding.     

The efficacy of a single chain recombinant equine luteinizing hormone (reLH) in mares: induction of ovulation, hormone profiles, and inter-ovulatory intervals

The objectives of this study were to determine the efficacy of recombinant equine luteinizing hormone (reLH) in shortening the time to ovulation in cycling mares and to determine the effects of treatment on endogenous hormones and inter-ovulatory intervals. In study 1, mares of light horse breeds (3–20 years) were treated with either a vehicle, various doses of reLH, or human chorionic gonadotropin (hCG). Cycling mares were examined by palpation and ultrasound per rectum daily or every 12 h from the time of treatment to ovulation. In studies 2 and 3, jugular blood samples were collected daily or every 12 h from the time of treatment to ovulation for analysis of LH, follicle stimulating hormone (FSH), estradiol-17β (E₂), and progesterone (P₄) by radioimmunoassays (RIA). Increasing doses of reLH (0.3, 0.6, 0.75, and 0.9 mg) showed increasing effectiveness at inducing ovulation within 48 h of treatment. Treatments with the 0.75 and 0.9 mg doses of reLH resulted in 90% and 80% ovulation rates, which were similar to hCG treatment (85.7%). Except for the early rise in LH after treatment with 0.5, 0.65, and 1.0 mg of reLH, hormone profiles appeared to be similar between control and treated cycles. Inter-ovulatory intervals were similar between control and treatment cycles. In conclusion, reLH is a reliable and effective ovulatory agent that does not significantly alter endogenous hormone profiles or affect inter-ovulatory intervals.     

利用重组自交系(RILs)群体进行质量数量性状的遗传分析遗传模型和小麦产量性状遗传

本文提出了利用RILs群体进行质量数量性状分析的4个遗传模型及统计分析方法.其中,模型Ⅰ、Ⅱ、Ⅲ、Ⅳ分别表示性状受1对主基因、2对独立的主基因、2对AABB×aabb型连锁的主基因、2对AAbb×aaBB型连锁的主基因与微效基因共同控制.可估算出主基因的加性效应(dma,dmb)、互作效应(im)、遗传方差(σ2m)和连锁图距(R),以及微效基因     

玉米作图群体主要穗部性状的研究

本研究调查了玉米自交系Mo17、黄早4及由这两个材料构建的F9代重组自交系(Recombinantinbred line,RIL)群体的穗粗、穗长、轴粗和穗重共4个穗部性状,对这4个性状在群体中的表现用SPSS11.5软件作了描述性统计和相关分析,并构建了频数分布图。该结果为控制这4个性状的数量性状位点(Quanti-tative trait locus,QTL)的作图研究提供了田间表型数据。     

表达鹅细小病毒VP3基因重组腺病毒的构建与鉴定

[目的]构建表达鹅细小病毒 （GPV） VP3基因的重组腺病毒,为机体免疫试验和免疫效果评价奠定基础.[方法]以重组质粒pcDNA-VP3为模板,以GPV-VP3为目的基因,构建GPV-VP3重组腺病毒载体,通过转染获得能稳定表达GPV-VP3基因的重组腺病毒,通过IFA和Western-blot检测GPV-VP3基因的表达情况.[结果]扩增到的GPV-VP3基因全长为1 605 bp,线性化的重组腺病毒穿梭质粒pCR259-VP3能在QBI-HEK293细胞中瞬时表达GPV-VP3基因,表达蛋白的分子量约为60Ku.[结论]该重组腺病毒的构建将为GPV新型疫苗研发和后期体内试验奠定基础.