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1.
低能离子束介导水稻遗传转化的研究进展   总被引:2,自引:0,他引:2  
介绍了低能离子束介导遗传转化的原理,比较了离子束介导法与其他遗传转化方法的优缺点,重点对离子束介导水稻遗传转化的最新研究进展进行了评述,同时提出了问题和展望.  相似文献   

2.
建立低能离子束介导小麦转基因方法并获得转GUS基因植株   总被引:42,自引:0,他引:42  
研究了注入离子种类、能量、剂量等参数对于低能离子束介导的遗传转化的影响,建立了适于小麦成熟胚转化的组培条件和筛选程序。以携带GUS基因的质粒为供体,进行了报告基因转化研究。分子生物学证据表明GUS基因已整合到小麦基因组中。3个小麦品种的抗性愈伤转化率分别为9.5%、10.8%、11.2%,再生植株转化率分别为1.4%、3.4%、1.7%,首次证明了离子束介导小麦遗传转化是可行的,为离子束介导小麦遗  相似文献   

3.
《生物技术通报》2005,(4):81-82
植物特别是高等植物在漫长的历史形成过程中所具有的遗传保守性一般很不易打破,但用生物技术方法,目前可以打破,且可创造新的突变体和新物种,由此可达此目的。例如郑州大学离子束生物工程实验室的卞坡博士和华中农业大学食品科技学院的谈重芳先生等利用离子束介导外源全DNA转化拟南芥菜的诱变效应就是例证之一。  相似文献   

4.
根癌农杆菌介导法(Agrohacterium mediated transformation)和基因枪轰击法( particle bombardment transformation)是植物遗传转化的主要方法。两种方法各有优缺点.农杆菌介导法是一种天然的植物遗传转化系统,外源基因在转基因植物中的拷贝数低,遗传稳定性好;基因枪转化法不受材料基因型的限制。通过结合两种方法的优点,发展了3种农杆菌介导和基因枪轰击法相结合的遗传转化方法,分别为农杆枪法、基因枪轰击/农杆菌感染法、金粉或钨粉包裹菌体细胞作为微弹轰击法。对3种结合转化方法的技术途径、原理、转化受体及研究进展等方面进行了综述。  相似文献   

5.
在离子束介导外源全DNA转化中,外源DNA包含有供体的所有遗传信息,对受体的影响是多方面的,因此转化效果无法用统一的转化率来衡量,这也增加了转化过程中最佳转化条件的选择难度。本论文的主要工作是根据不同离子剂量介导外源全DNA转化的拟南芥菜在遗传和生理上的变化选择合适的转化剂量。剂量实验结果表明:拟南芥菜种子的剂量-存活率曲线为"马鞍型",在"鞍桥"上选择0.5×1017ions/cm2、1.5×1017ions/cm2和2.5×1017ions/cm2三个有代表性的剂量介导薄荷全DNA转化拟南芥菜,三个转化当代群体的出芽、成苗长势以及表型变异情况有明显的不同,综合分析表明:1.5×1017ions/cm2为离子束介导外源全DNA转化拟南芥菜的合适剂量。  相似文献   

6.
高兴喜  杨谦 《微生物学报》2005,45(1):129-131
根癌农杆菌介导的遗传转化系统是植物基因工程常用方法,目前已将这一转化系统应用到酵母、丝状真菌以及人类细胞的转化。利用这一转化系统,成功地实现了丝状真菌球毛壳菌(Chaetomium globosum)的遗传转化,转化率约为60~180个转化子/10.7个孢子 。通过对转化子的PCR检测和Southern 杂交分析表明,TDNA已整合进毛壳菌基因组中,而且在所检测的转化子中都是以单拷贝的形式整合,转化子都能够稳定遗传。根癌农杆菌介导的遗传转化具有转化率高、低拷贝、遗传稳定、操作简便等优点,因此有可能成为丝状真菌遗传转化和功能基因组研究的有力工具。  相似文献   

7.
高效遗传转化技术是植物重要性状功能基因鉴定的前提和转基因育种的基础。随着纳米生物技术的发展,以纳米载体介导的植物转基因技术已显示出巨大的应用潜力。综述了国内外应用于植物纳米载体的类型、与外源基因的结合方式以及传输细胞的原理,重点阐述了影响纳米基因载体性能与转化效率的重要因素,以及纳米载体介导外源基因转化植物细胞的方法,分析了纳米载体介导法与其他转基因方法的特点和优势,并提出纳米载体介导的转化技术应加强稳定遗传转化、基因编辑与植物原位转化等方面探索研究,旨为植物遗传转化技术和方法提供新的思路。  相似文献   

8.
根癌农杆菌介导真菌遗传转化的研究进展   总被引:20,自引:0,他引:20  
根癌农杆菌介导的真菌遗传转化是近年来发展的一种新方法 ,与其它方法相比 ,该方法具有操作简便、转化效率高和易得到稳定转化子等特点。目前 ,在根癌农杆菌介导下已实现了多个属种真菌的遗传转化 ,显示出良好的应用前景。综述了根癌农杆菌介导真菌遗传转化的转化机理和T DNA在真菌细胞中的存在方式等方面的研究结果 ,并展望这一方法的应用前景。  相似文献   

9.
随着农杆菌介导遗传转化过程中农杆菌一方转化机理的阐明,人们现在已经将目光转向了参与在农杆菌介导遗传转化过程中的植物因子。应用拟南芥突变体分析,酵母双杂交和cDNA的扩增片段长度多态性等技术人们已经部分阐明了参与在农杆菌向植物细胞的附着,TDNA的加工和转运,T链复合体向细胞核的转运,TDNA的整合等过程中的植物因子,并对植物基因对于农杆菌侵染的响应有了初步的了解。这些结果的获得不仅有助于人们加深对农杆菌介导遗传转化机理的认识而且有助于进一步扩大农杆菌介导遗传转化这一技术的应用范围 。  相似文献   

10.
杨天佑  卞坡  谈重芳  吴健  秦广雍 《激光生物学报》2003,12(5):398-400,F003
离子束作为一种介导DNA转化技术逐渐得到了越来越广泛的应用,并且在促进DNA转化上存在很大优势。育种过程中任意引物PCR技术通过对诱变前后整个基因组的分析,在分子水平上提供了与宏观优势性状密切联系的特异性DNA片段,这些差异DNA片段在育种过程中是很重要的。借助离子束将这些片段转化于生物体细胞继续进行选育,可能对稳定品系及保持优良性状起到很大的作用。本文探讨了利用离子束转化RAPD差异片段的可行性,优势及其在育种中的前景。  相似文献   

11.
低能离子束在生物技术中的应用研究   总被引:45,自引:0,他引:45       下载免费PDF全文
自从发现离子注入生物效应后,低能离子与生物体系相互作用研究在我国率先兴起,并很快投入应用。简要介绍低能离子注入生物效应的机理研究和应用研究的进展状况,并展望未来 。  相似文献   

12.
低能氮离子注入大肠杆菌诱发的生物学效应研究   总被引:2,自引:0,他引:2  
研究离子注入的诱变和导入外源DNA的生物学效应,用低能氮离子注入处理大肠杆菌野生型菌株MC4100A,将含水母绿色荧光蛋白基因的质粒导入细胞中。实验结果表明,在一些转化子中绿色荧光蛋白的表达或折叠受到了影响,一些转化子丧失了分裂后分离能力,且细胞膜有一定程度的损伤,为进一步研究微生物细胞分裂和蛋白质折叠机理提供了菌株。故低能氮离子注入对微生物细胞的诱变效应和导入外源DNA效应的有机结合将使离子注入技术在生物学基础研究中有着广泛的应用前景。  相似文献   

13.
A cytogenetic study of the activity of the embryonic genome in ferret (Mustela putorius) blastocysts during ∼6 days after their transition from the oviduct to the uterus has been carried out. It has been found that the prolongation in the preimplantation period in the ferret is not accompanied by inhibition of mitosis or activity in nucleolus organizing regions of inner cell mass cells as occurs in species having an obligatory delay of implantation (obligate embryonic diapause). Amitosis of trophoblast cells starts at the periimplantation stage as in other species that do not have obligate diapause. The data obtained are consistent with the hypothesis that the obligatory stage of delayed implantation might occur in some mammals in different taxonomic groups as a result of chromosome mutations affecting the genetic control of the chronology of events (timing) of embryogenesis. Consequently the characteristics of delayed implantation should be different in different species.  相似文献   

14.
Infertility and spontaneous pregnancy losses are an enduring problem to women's health. The establishment of pregnancy depends on successful implantation, where a complex series of interactions occurs between the heterogeneous cell types of the uterus and blastocyst. Although a number of genes are implicated in embryo-uterine interactions during implantation, genetic evidence suggests that only a small number of them are critical to this process. To obtain a global view and identify novel pathways of implantation, we used a dual screening strategy to analyze the expression of nearly 10,000 mouse genes by microarray analysis. Comparison of implantation and interimplantation sites by a conservative statistical approach revealed 36 up-regulated genes and 27 down-regulated genes at the implantation site. We also compared the uterine gene expression profile of progesterone-treated, delayed implanting mice to that of mice in which delayed implantation was terminated by estrogen. The results show up-regulation of 128 genes and down-regulation of 101 genes after termination of the delayed implantation. A combined analysis of these experiments showed specific up-regulation of 27 genes both at the implantation site and during uterine activation, representing a broad diversity of molecular functions. In contrast, the majority of genes that were decreased in the combined analysis were related to host immunity or the immune response, suggesting the importance of these genes in regulating the uterine environment for the implanting blastocyst. Collectively, we identified genes with recognized roles in implantation, genes with potential roles in this process, and genes whose functions have yet to be defined in this event. The identification of unique genetic markers for the onset of implantation signifies that genome-wide analysis coupled with functional assays is a promising approach to resolve the molecular pathways required for successful implantation.  相似文献   

15.
16.
Bmp and Fgf signaling are essential for liver specification in zebrafish   总被引:2,自引:0,他引:2  
Based on data from in vitro tissue explant and ex vivo cell/bead implantation experiments, Bmp and Fgf signaling have been proposed to regulate hepatic specification. However, genetic evidence for this hypothesis has been lacking. Here, we provide in vivo genetic evidence that Bmp and Fgf signaling are essential for hepatic specification. We utilized transgenic zebrafish that overexpress dominant-negative forms of Bmp or Fgf receptors following heat-shock induction. These transgenes allow one to bypass the early embryonic requirements for Bmp and Fgf signaling, and also to completely block Bmp or Fgf signaling. We found that the expression of hhex and prox1, the earliest liver markers in zebrafish, was severely reduced in the liver region when Bmp or Fgf signaling was blocked just before hepatic specification. However, hhex and prox1 expression in adjacent endodermal and mesodermal tissues appeared unaffected by these manipulations. Additional genetic studies indicate that the endoderm maintains competence for Bmp-mediated hepatogenesis over an extended window of embryonic development. Altogether, these data provide the first genetic evidence that Bmp and Fgf signaling are essential for hepatic specification, and suggest that endodermal cells remain competent to differentiate into hepatocytes for longer than anticipated.  相似文献   

17.
离子束在生命科学中的应用   总被引:11,自引:4,他引:7  
吴丽芳  李红 《激光生物学报》1999,8(4):299-304,311
低能离子在生命科学中应用是道德在中国兴起的1986年,中国的科技工作者研究了低能哼子对生物的诱变效应,并成功地将其应用于植物育种。现在,离子注入技术已被应用于生物改良、生命起源和进化以及环境辐射生物学效应等多个理论和应用研究领域。其中,在植物遗传转化、创造新种质资源方面的成果尤其为突出:通过离子束介导转化法获得了高蛋白小麦、高光效水稻,抗病小麦、水稻、棉花新品质。通过诱变技术获得了多穗型玉米、无融  相似文献   

18.
Gene sequences in human oocytes were studied to investigate the possibility of diagnosing inherited or sporadic genetic disease before implantation after in vitro fertilisation. By specific amplification the possibility of analysing the DNA from single human oocytes for a specific gene was shown, and genotypes for markers closely linked to cystic fibrosis and Duchenne muscular dystrophy were determined. Single oocytes were used to approximate the total amount of DNA present in a single cell taken for biopsy from a 4-16 cell blastocyst. With a new technique for specific DNA amplification, the polymerase chain reaction, these data can be obtained within several hours of cell isolation. Extreme care must be taken to avoid any contamination of the sample with DNA from other sources. With this technique genotyping for single gene disorders is feasible with an accuracy and on a time scale that would allow implantation of the zygote after in vitro fertilisation without freezing.  相似文献   

19.
There are reports of adverse effects of cannabinoids on pregnancy outcome including retarded embryo development and pregnancy failure. Thus, discoveries of endogenous cannabinoid-like lipid mediators and cannabinoid receptors raise questions about their pathophysiological roles during normal pregnancy. We previously reported that anandamide, an endogenously produced arachidonate derivative (endocannabinoid), is synthesized in the female reproductive tracts, and it acts on cannabinoid receptors expressed on the cell surface of the embryo to regulate the preimplantation embryo development and implantation in mice. This review presents genetic, molecular, physiological and pharmacological evidence that the levels of uterine anandamide and blastocyst CB1 cannabinoid receptors are coordinately regulated to synchronize preimplantation development and uterine receptivity for implantation in mice.  相似文献   

20.
The results of a variety of studies on the genetic and immunological aspects of reproduction can be integrated into a hypothesis about the factors that regulate implantation and development and that may also cause an increased susceptibility to cancer. The primary condition for successful reproduction is genetic compatibility between the mating partners: there must be no recessive lethal genes that could act alone or epistatically to cause embryonic or fetal death. Such recessive lethal genes have been identified in the mouse (t-haplotypes) and in the rat (grc), and there is some evidence that they also exist in humans. Immunological factors may modulate the implantation of the fertilized ovum under some cireumstances after the genetic condition has been met. The same genetic factors that affect development may also affect susceptibility to cancer. This part of the hypothesis is supported by a number of clinical correlations between congenital defects and a higher incidence of cancer and by the demonstration of an increased susceptibility to the effects of chemical carcinogens in rats carrying the grc.  相似文献   

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