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81.
Shukuro Araki Shigehiro Yi Tatsufumi Murakami Susumu Watanabe Shinichi Ikegawa Kiyoshi Takahashi Ken-ichi Yamarnura 《Molecular neurobiology》1994,8(1):15-23
To analyze the pathologic processes of amyloid deposition in type I familial amyloidotic polyneuropathy (FAP), mice were made
transgenic by introducing the human mutant transthyretin (TTR) gene(MT-hMet 30). An inbred strain of mouse, C57 BL/6, was
chosen. Transgenic mice were killed using ether anesthesia at 3-mo intervals up to 24 mo after birth. In these transgenic
mice, amyloid deposition started in the gastrointestinal tract, cardiovascular system, and kidneys and extended to various
other organs and tissues with advancing age. The pattern of amyloid deposition was similar to that observed in human autopsy
cases of FAP, except for its absence in the choroid plexus and in the peripheral and autonomic nervous systems.
We extracted the amyloid fibrils from kidneys of these mice with a human mutant TTR gene and analyzed them immunochemically
and electronmicroscopically. Deposited amyloid was shown to be composed of human mutant TTR and mouse serum amyloid P component.
Amyloid fibril from transgenic mice was morphologically and immunohistochemically similar to that of human FAP.
The most striking pathologic feature of the transgenic mice was the absence of amyloid deposition in the peripheral and autonomic
nervous tissues. Thus, other intrinsic factors may be involved in amyloid deposition in the nervous tissues of human FAP. 相似文献
82.
Ming-Tsair Chan Hsin-Hsiung Chang Shin-Lon Ho Wu-Fu Tong Su-May Yu 《Plant molecular biology》1993,22(3):491-506
We have successfully transferred and expressed a reporter gene driven by an -amylase promoter in a japonica type of rice (Oryza sativa L. cv. Tainung 62) using the Agrobacterium-mediated gene transfer system. Immature rice embryos (10–12 days after anthesis) were infected with an Agrobacterium strain carrying a plasmid containing chimeric genes of -glucuronidase (uidA) and neomycin phosphotransferase (nptII). Co-incubation of potato suspension culture (PSC) with the Agrobacterium inoculum significantly improved the transformation efficiency of rice. The uidA and nptII genes, which are under the control of promoters of a rice -amylase gene (Amy8) and Agrobacterium nopaline synthase gene (nos), respectively, were both expressed in G418-resistant calli and transgenic plants. Integration of foreign genes into the genomes of transgenic plants was confirmed by Southern blot analysis. Histochemical localization of GUS activity in one transgenic plant (R0) revealed that the rice -amylase promoter functions in all cell types of the mature leaves, stems, sheaths and roots, but not in the very young leaves. This transgenic plant grew more slowly and produced less seeds than the wild-type plant, but its R1 and R2 progenies grew normally and produced as much seeds as the wild-type plant. Inheritance of foreign genes to the progenies was also confirmed by Southern blot analysis. These data demonstrate successful gene transfer and sexual inheritance of the chimeric genes. 相似文献
83.
84.
The expression of an Arabidopsis acyl carrier protein (ACP) gene promoter has been examined in transgenic tobacco plants by linking it to the reporter gene -glucuronidase (GUS). Fluorometric analysis showed that the ACP gene promoter was most active in developing seeds. Expression was also high in roots, but significantly lower in young leaves and downregulated upon their maturation. Etiolated and light-grown seedlings showed the same level of GUS activity, indicating that this promoter is not tightly regulated by light. Histochemical studies revealed that expression was usually highest in apical/ meristematic zones of vegetative tissues. Young flowers (ca. 1 cm in length) showed GUS staining in nearly all cell types, however, cell-specific patterns emerged in more mature flowers. The ACP gene promoter was active in the stigma and transmitting tissue of the style, as well as in the tapetum of the anther, developing pollen, and ovules. The results provide evidence that this ACP gene is regulated in a complex manner and is responsive to the array of signals which accompany cell differentiation, and a demand for fatty acids and lipids, during organogenesis. 相似文献
85.
Tomatoes (Lycopersicon esculentum Mill cv. Ailsa Craig) were transformed with a gene construct having 244 bp of the 5 end of a polygalacturonase (PG) cDNA, coding for a 71 amino acid N-terminal extension to the mature protein, fused to 1320 bp of a pectinesterase (PE) cDNA encoding the full sequence of the mature PE protein. This chimaeric gene was inserted in a sense orientation between a CaMV 35S promoter and terminator for constitutive expression. In transformed tomato plants expression of the endogenous PG and PE genes in the fruit was inhibited; there was little or no observable PG and PE mRNA and a substantial reduction in the level of PG and PE enzyme activity. The transgene was expressed in the leaves of the transformed plants as demonstrated by the accumulation of mRNA, but no protein product could be identified. However, no transgene mRNA or protein were observed in the transgenic fruit.This paper represents the first report of the down-regulation of two non-homologous endogenous genes using a single gene construct. A sense gene construct was responsible for these effects. These findings are discussed in relation to possible mechanisms of action of co-suppression. 相似文献
86.
Direct DNA transfer methods based on particle bombardment have revolutionized plant genetic engineering. Major agronomic crops
previously considered recalcitrant to gene transfer have been engineered using variations of this technology. In many cases
variety-independent and efficient transformation methods have been developed enabling application of molecular biology techniques
to crop improvement. The focus of this article is the development and performance of electric discharge particle bombardment
(ACCELL™) technology. Unique advantages of this methodology compared to alternative propulsion technologies are discussed
in terms of the range of species and genotypes that have been engineered, and the high transformation frequencies for major
agronomic crops that enabled the technology to move from the R&D phase to commercialization.
Creation of transgenic soybeans, cotton, and rice will be used as examples to illustrate the development of variety-independent
and efficient gene transfer methods for most of the major agronomic crops. To our knowledge, no other gene transfer method
based on particle bombardment has resulted in variety-independent and practical generation of large numbers of independently-derived
crop plants. ACCELL™ technology is currently being utilized for the routine transfer of valuable genes into elite germplasm
of soybean, cotton, bean, rice, corn, peanut and woody species. 相似文献
87.
喹诺酮类药物抗乙型肝炎病毒体外实验研究 总被引:4,自引:0,他引:4
本文以2.2.15细胞株为模型,以HBsAg、HBeAg、HBVDNA、细胞存活率为观察指标,综合评价了喹诺酮类药物吡哌酸(PipemidicAcid)、氟哌酸(Norfloxacin)、环丙氟哌酸(Ciproflosxacin)、氟嗪酸(Ofloxacin)体外抗HBV效果。结果表明:吡哌酸、氟哌酸、环丙氟哌酸、氟嗪酸对HBsAg、HBeAg50%抑制浓度(ID_(50))分别为11μg/ml、64μg/ml、93μg/ml、105μg/ml和199μg/ml、111μg/ml、24μg/ml、217μg/ml,细胞存活率为50%时的药物浓度(CD_(50))分别为219μg/ml、90μg/ml、181μg/ml、169μg/ml,在所选定的用药浓度范围内不同程度抑制培养上清液及细胞内HBVDNA及其复制中间体的产生。尤其对超螺旋结构DNA(scDNA)有不完全抑制作用。 相似文献
88.
P. J. Hatt M. Moriniere H. Oberlander P. Porcheron 《In vitro cellular & developmental biology. Animal》1994,30(10):717-720
Summary During postembryonic development of insects, molting cycles affect epidermal cells with alternate periods of proliferation
and differentiation. Cells of the cell line established from imaginal discs of the Indian meal moth (IAL-PID2) differentiate
under the action of the molting hormone, 20-hydroxyecdysone, in a manner that is meaningful in terms of the development of
the tissue from which they were derived. In particular, the hormone caused an accumulation of the cells in the G2 phase of
their cycle and induced the formation of epithelial-like aggregates and the synthesis of specific proteoglycans. Recent discovery
of members of the insulin superfamily in insects and the role of growth factors played by this family of molecules in vertebrates
led us to check for their potential effects on IAL-PID2 cell cycle regulation. On the one hand, our results showed that insulin
was involved in partial resumption of the cell cycle after an arrest caused by serum deprivation, but that other growth factors
present in fetal calf serum were needed for full completion of mitosis. On the other hand, the cytostatic effect of 20-hydroxyecdysone
was reversible, and, prior exposure of the cells to the hormone allowed the cells to complete one cell cycle in serum-free
medium. These results suggest that the production of autocrine growth factors induced by ecdysteroids could circumvent the
absence of serum. This cell culture model provides potential for further study of interactions between ecdysteroids and growth
factor homologs during differentiation of insect epidermal cells. 相似文献
89.