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71.
Role of viral antigens in destructive cellular immune responses to adenovirus vector-transduced cells in mouse lungs. 总被引:8,自引:3,他引:5 下载免费PDF全文
Adenoviruses missing E1 have been used as gene delivery vectors to the lungs for the treatment of cystic fibrosis. Transient expression of the recombinant gene and the development of inflammation have been two major limitations to the application of first-generation recombinant adenoviruses for gene therapy. Studies with mouse models of liver- and lung-directed gene therapy suggested that CD8+ cytotoxic T lymphocytes (CTLs) are effectors that contribute to extinction of transgene expression. The precise antigens responsible for activation of CTLs have not been identified. In this study, we examine the relative contributions of viral proteins versus the transgene product to the activation of CTLs which eliminate transgene-containing cells in mouse lungs. Instillation of a lacZ-expressing virus into the lungs of C57BL/6 mice elicited CTL responses to both viral proteins and the transgene product, beta-galactosidase, which collectively contribute to loss of trans-gene expression in mouse airways. Similar results were obtained in two experimental models in which the animals should be tolerant to the transgene, i.e., lacZ virus delivered to an animal transgenic for lacZ and a virus expressing the liver-specific enzyme ornithine transcarbamylase administered to the lungs of various strains of immune-competent mice. These data confirm the hypothesis that CTLs specific for viral antigens contribute to the problem of transgene instability in mouse lungs and indicate that CTLs specific for transgene product alone cannot account for the observed problem. 相似文献
72.
†Kelly J. Conn Michael Pietropaolo ‡Shyr-Te Ju † Carmela R. Abraham 《Journal of neurochemistry》1996,66(5):2011-2018
Abstract: Alzheimer's disease is characterized neuropathologically by the presence of neuritic and amyloid plaques, vascular amyloid, and neurofibrillary tangles in specific brain areas. The main constituent of amyloid deposits is amyloid β protein, a 40–42 amino acid proteolytic product of the amyloid β-precursor protein. In our search for proteases that can generate the N-terminus of amyloid β protein (β-secretases), we discovered a thiol-dependent metalloprotease that was identified, by peptide sequencing, as metalloendopeptidase EC 3.4.24.15. In vitro, the metalloprotease cleaves the methionine-aspartic acid bond in a 10 amino acid synthetic peptide, indicating that it could generate the N-terminus of amyloid β protein, and generates amyloidogenic fragments from full-length recombinant amyloid β-precursor protein. Mouse monoclonal antibodies produced against a unique synthetic peptide from the metalloprotease labeled various monkey tissues as detected by western blots and immunohistochemistry. Unexpectedly, two monoclonal antibodies, IVD6 and IIIF3, immunolabeled strongly intracellular neurofibrillary tangles, neurites of senile plaques, and neuropil threads, but not "ghost" tangles or amyloid in sections taken from Alzheimer's disease brain. This finding provides further evidence for the metalloprotease's relevance to Alzheimer's disease pathology, although the connection between tangle staining and the formation of amyloid β protein remains to be elucidated. 相似文献
73.
Maria de Fatima Bonaldo Pierre Jelenc Long Su Lee Lawton M. -T. Wu Dorothy Warburton Marcelo Bento Soares 《Human genetics》1996,97(4):441-452
A study was conducted on the feasibility of isolating genes and pseudogenes that map to chromosome 13 by a hybridization-based approach using a 13-specific library and pools of repeat-free cDNA clones. Five pairs of cDNA and chromosome 13 genomic clones were identified and characterized. Partial or full-length sequence was derived from all cDNAs, and database searches were performed for putative gene identification. Partial sequence was also obtained from the chromosome 13 genomic clones for comparison with those of the hybridizing cDNAs. As a result of these analyses we identified three genes, a putative homologue of a porcine mRNA encoding an unidentified hepatic protein, a putative homologue of a yeast integral membrane protein, and a gene for a translationally controlled tumor protein, and two processed pseudogenes, ribosomal proteins L23a and S3a. The latter was formerly identified as the v-fos transformation effector gene, Fte-1, and recently cited as a possible candidate for the BRCA2 gene on chromosome 13. All genes and pseudogenes were localized to cytogenetic bands by in situ hybridization of metaphase chromosomes with probes derived from the chromosome 13 genomic clones. 相似文献
74.
棕色固氮菌缺失nifZ基因的突变种固氮酶MoFe蛋白的纯化和性质 总被引:1,自引:0,他引:1
采用 52℃下加热 6 min,后经 DEAE- 52、Sephacryls S- 2 0 0和 Q- Sepharose等柱层析方法 ,分离纯化了棕色固氮菌 (Azotobacter vinelandii)缺失 nif Z基因突变种固氮酶 Mo Fe(Δnif Z Mo Fe)蛋白 ,其纯度达到电泳纯。Δnif Z Mo Fe蛋白的固氮活性为 2 83nmol C2 H2 还原 / (min·mg蛋白 ) ,远低于野生种 Mo Fe蛋白。Δnif Z Mo Fe蛋白对氧更敏感 ;热稳定性略低于野生种。Δnif Z Mo Fe蛋白的可见光吸收光谱与野生种 Mo Fe蛋白极为相似。其圆二色谱和磁圆二色谱在 450~ 550 nm与野生种 Mo Fe蛋白显著不同 ,表明其 P- cluster及其周围环境与野生种 Mo Fe蛋白有所差异。这亦可能是造成缺失 nif Z突变种 Mo Fe蛋白固氮活性低的原因。 相似文献
75.
菜用大豆产量相关性状的遗传分析 总被引:7,自引:0,他引:7
对 19个菜用大豆品种与产量有关的10个农艺性状进行遗传分析的结果表明,生育期和主茎节数遗传力偏高;单株荚数、分枝数遗传力偏低;单株产量、单株荚数的遗传变异系数很大,其遗传进度的值也较大;生育期的遗传变异系数小, 遗传进度也小,遗传相关分析结果表明,产量与生育期、单株结荚数相关关系密切。菜用大豆遗传参数分析结果与前人对食用大豆的研究结果趋势一致。 相似文献
76.
懒猴属的核糖体DNA变异及其种间分化关系 总被引:5,自引:2,他引:3
用15种限制性内切酶和人28S、18SrDNA探针构建了懒猴属各物种核糖体DNA重复单位的限制性内切酶图谱。在进化速率较高的非转录间隔区,在大、中、小懒猴中分别定位了23、24、24个酶切位点。大懒猴与中懒猴有12个位点不同,与小懒猴有14个位点不同,而中、小懒猴间则只有一个位点的差异。经过计算,大懒猴与中懒猴的遗传距离值为12.65%,与小懒猴的差异为14.24%,中、小懒猴间的差异则仅为0.7 相似文献
77.
云南姬鼠的蛋白多态性及其遗传分化关系 总被引:3,自引:0,他引:3
本文采用蛋白电泳技术对来源于云南省若干地区的姬鼠属(Apodemus)的3种姬鼠──高山姬鼠(A.chevrieri)8只,中华姬鼠(A.draco)3只和大耳姬鼠(A.latronum)1只,以及作为外群的同科的绒鼠属的大绒鼠(Hapalomysdelalori)3只进行了分析。共检测遗传座位27个,发现21个座位存在多态性。根据蛋白多态的数据对研究对象进行遗传分化关系的探讨,用系统分析软件PHYLIP计算它们之间的分化关系,得到了一棵无根系统树。结果表明,作为外群的大绒鼠明显不同于其它3种姬鼠而聚在最外面。8只高山姬鼠个体汇聚成独立的一支,中华姬鼠的3个个体也聚成一支,但大耳姬鼠却聚在中华姬鼠一支中,因此我们认为大耳姬鼠同中华姬鼠的分化时间可能比较晚近。 相似文献
78.
79.