共查询到19条相似文献，搜索用时 218 毫秒
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真核生物mRNA差异显示技术的创立及其对该技术的一系列改进,为研究胚胎发育过程中有关基因的差异表达,以及有关基因的分子克隆提供了有效工具.文章概述了差显技术的原理及其在动物早期胚胎发育基因方面的研究进展. 相似文献
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mRNA差异显示技术及其在动物科学研究中的应用 总被引：5，自引：1，他引：5
基因表达的变化是调控细胞生命活动过程的核心机制,分析不同细胞或同类细胞在不同发育阶段、不同生理状态下基因的表达状况,可以为研究生命活动过程提供重要信息。mRNA差异显示技术(differentialdisplay,DD)是目前应用比较广泛的在mRNA水平上检测基因表达的方法之一。本文介绍了mRNA差异显示技术原理及其优缺点,对包括引物设计改进、反应条件的优化、差异条带显示方法改进和降低假阳性策略等在内的技术改进进行了概述,对mRNA差异显示技术在动物生理学和病理学、动物胚胎、个体发育、动物遗传育种、动物营养研究中的应用作了概括介绍。 相似文献
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转基因表达的调控方法 总被引：4，自引：0，他引：4
转基因动物技术作为一种研究基因功能的技术体系 ,在生命科学研究领域有着广泛的应用前景。有效地使精确的遗传基因修饰在动物体内得到表达并实现世代间的传递是转基因技术的关键。近年来由于转座子、逆转录病毒的运用以及采用加入或去除某些基因的体细胞的克隆技术的发展 ,不同物种转基因的培育方法日趋简便。 Cre-L ox P系统越来越多地用于从基因组中去除特定的序列或靶向整合外源DNA。四环素等系统已被证实可获得确切的转基因表达。具有反式显性阴性效应的与 DNA形成三链螺旋的 RNA、反义 RNA(包括 :含 RNA干预和核酶的双链 RNA)以及蛋白质的表达均被证实可特异性地抑制宿主或病毒基因的表达。文章综述了转基因的概念、表达及调控转基因表达的常用方法 相似文献
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银染mRNA差异显示法克隆柔嫩艾美耳球虫(Eimeria tenella)抗药性相关基因 总被引：5，自引：0，他引：5
以柔嫩艾美耳球虫（Eimeria tenella）敏感株、地克珠利抗药株和马杜拉霉素抗药株孢子化卵囊为材料，用银染mRNA差异显示方法筛选和克隆与球虫抗药性相关的基因。首先提取这3个虫株孢子化卵囊的总RNA为模板．用Oligo（dT）12GG为锚定引物和2个10碱基随机引物组合进行RTPCR，产物经变性聚丙烯酰胺凝胶电泳后银染。分别切取5务差异条带，进行2次PCR扩增，产物回收后与PGEM—T—easy Vector连接转化。经PCR鉴定正确后，再进行斑点杂交试验、序列分析和同源性比较。结果发现，地克珠利抗药株和马杜拉霉素抗药株分离的差异片段中都有2个cDNA片段（可能为新的基因片段），这为克隆全长cDNA和探索球虫抗药性产生的分子机理奠定了一定的基础。 相似文献
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Embryo transfer: its uses and recent developments 总被引：3，自引：0，他引：3
J M Sreenan 《The Veterinary record》1988,122(26):624-629
The technique of embryo transfer is now routinely employed in several species and with several objectives. In laboratory animals the technique is used mainly in investigations of reproductive biology, whereas in human beings it is used to overcome specific forms of impaired fertility. Because embryo transfer combined with superovulation of the donor can significantly increase the female reproductive rate its greatest application to date has been in farm animals, in which it is widely used in both research and commercial production. Within the past few years there have been many advances in the techniques used in farm animals, particularly in the area of embryo manipulation. The supply of embryos can now be increased by repeated superovulation and by embryo bisection and there has been significant progress in in vitro fertilisation technology. Deep freeze (-196 degrees C) storage of embryos is now routine and may be combined with direct one-step thawing and removal of cryoprotectant. This technique allows the routine non-surgical transfer of embryos in the field. There is also evidence that a routine non-traumatic procedure for sexing embryos may soon be developed. Identical multiplets or clones have been produced by the microdissection of embryos and more recently by the transfer of nuclei from embryos into unfertilized oocytes. Transgenic animals have been produced by the microinjection of recombinant DNA into one of the pronuclei of single-cell unfertilized ova. 相似文献
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噬菌体肽库技术筛选抗PRRSV肽及其应用 总被引：1，自引：0，他引：1
噬菌体展示肽库是一种被广泛用于抗原表位鉴定,结合蛋白筛选的技术。该试验利用噬菌体展示技术筛选与猪呼吸与繁殖障碍综合征病毒（PRRSV）ORF1B复制酶蛋白相互作用的蛋白,并进一步验证筛选蛋白的抗病毒作用。以表达纯化的PRRSV ORF1B蛋白CTD包被高亲和性96孔板作为靶蛋白,应用T7噬菌体展示技术对随机12肽库cDNA文库进行筛选,并分析测序筛选的克隆。将筛选出的克隆序列合成后,在体外验证合成多肽的抗PRRSV效果。结果表明,在4轮的噬菌体筛选后,共得到87个阳性克隆,经测序鉴定出11个筛选的12肽,并通过体外抗病毒试验得到P10是一个具有高抗病毒活性的12肽。试验结果为PRRSV的抗病毒研究奠定了基础。 相似文献
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Many Newcastle disease virus strains are composed of several biotypes which coexist in the wild and in laboratory cultures. We have studied some of the phenotypic and genotypic properties of 6 virus clones from the coexisting biotypes of the Hickman strain of Newcastle disease virus. These clones were readily distinguishable from the parent virus strain and from each other by their RNA fingerprints. Fingerprints of the most virulent clones (Hi/LC, Hi/MC, and Hi/LR) contained 61% to 77% of the oligoribonucleotides present in the fingerprint of the Hickman parent strain. None of the clones killed chickens as rapidly as did the parent strain, although some clones killed embryonating eggs as rapidly as did the parent strain. 相似文献