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The functional activity of the promoter unit contained within the long terminal repeat (LTR) of bovine leukemia virus (BLV) was examined by monitoring transient expression of a heterologous gene placed under its control. Various cell lines were transfected with recombinant plasmids carrying the bacterial chloramphenicol acetyltransferase (CAT) gene coupled to the BLV LTR (pBL-cat). Transient expression of CAT activity directed by the BLV LTR was observed only in the established BLV-producer cell lines derived from fetal lamb kidney (FLK) cells and bat lung cells. The amount of CAT activity transiently expressed in FLK-BLV cells was decreased approximately tenfold by deletion of LTR sequences located within a region 100 to 170 nucleotides upstream of the RNA start site. Surprisingly, removal of the region 50 base pairs downstream of the RNA initiation site to the 3'-end of the LTR reduced the expression of CAT activity by 87 percent. The BLV LTR thus appears to be an unusual promoter unit, functioning in a cell type-specific manner and possessing sequences on both the 5' and 3' sides of the RNA start site that influence gene expression. 相似文献
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M. I. Gulyukin N. G. Kozyreva L. A. Ivanova T. V. Stepanova I. A. Gulyukina 《Russian Agricultural Sciences》2016,42(6):472-475
Nucleotide sequence of a section of the Env 105 gene of bovine leukemia provirus isolates, obtained from farm animals in different regions of Russia, was determined. Conducted phylogenetic analysis has allowed us to assess heterogeneity of the studied population of the bovine leukemia virus (BLV). Based on results of BLV genetic diversity, four virus’s genotypes—I, II, IV, and VII with dominating genotype IV—were detected. 相似文献
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参考BVDVVEDEVAC株的基因组序列及E2基因的测序结果设计一对引物,扩增出预期585 bp的目的片段。扩增产物克隆至pMD18-T载体,经酶切鉴定获得阳性重组质粒并对其进行测序。测序结果与参考序列VEDE-VAC株比较,二者的核苷酸同源性为100%,推导氨基酸同源性为100%。测序结果经同源性比较,克隆得到的基因与VEDEVAC株同源性最高。系统发生树分析推测E1基因与VEDEVAC株在进化上比较接近。将E1基因定向亚克隆到表达载体pGEX-6p-1中,对阳性重组质粒转化的大肠杆菌BL21进行诱导,E1基因获得了成功表达。 相似文献
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Cep164基因是参与纤毛形成的重要基因之一,为研究Cep164基因与CP110的关系,探讨CP110在Cep164调节纤毛形成中的作用,通过免疫荧光染色与Western印迹法确定Cep164基因缺失时CP110基因的表达情况,利用免疫共沉淀法确认Cep164与CP110蛋白相互作用的关系。结果表明,Cep164基因的缺失导致CP110表达增高,Cep164与CP110蛋白无直接相互作用关系,Cep164是通过间接方式调控CP110在细胞内表达,CP110降解失败可能为Cep164基因缺失导致纤毛生成障碍的原因之一。 相似文献
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克隆了牛的myf6基因并构建真核表达载体pIRES2-EGFP-myf6,用脂质体技术转染鲁西黄牛成纤维细胞,通过G418筛选出稳定转染的细胞株,利用Western印记、Real-time PCR技术检测myf6对成纤维细胞的影响。结果表明,与对照组相比,稳定转染后的成纤维细胞myf6蛋白和mRNA的表达量提高(P0.01),肌肉肌酸激酶基因mRNA表达量升高(P0.05),肌球蛋白轻链基因的mRNA表达量也提高(P0.01)。细胞形态观察显示转染后的成纤维细胞未融合为肌管。由此可知,牛myf6基因可以在成纤维细胞中表达并且有促进成纤维细胞向肌肉细胞分化的功能。 相似文献
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Two mechanisms for the extinction of gene expression in hybrid cells 总被引:12,自引:0,他引:12
When two different mammalian cell types are fused to generate a stable hybrid cell line, genes that are active in only one of the parents are frequently shut off, a phenomenon called extinction. In this study two distinct, complementary mechanisms for such extinction of growth hormone gene expression were identified. In hybrids formed by fusing fibroblasts to pituitary cells, pituitary-specific proteins that bind to the growth hormone promoter were absent. In addition, a negative regulatory element located near the rat growth hormone promoter was specifically activated. 相似文献
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根据GenBank中公布的牛BMAP-28基因mRNA序列设计引物,利用RT-PCR技术从牛骨髓总RNA中扩增出BMAP-28基因片段,将其克隆到pMD-18载体上,通过PCR、酶切和测序分析鉴定,获得重组克隆载体pMD18-T-BMAP28.以重组质粒为模板,扩增BMAP-28基因的去信号肽片段,转入原核表达载体PE... 相似文献
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Light pulses that shift rhythms induce gene expression in the suprachiasmatic nucleus 总被引:22,自引:0,他引:22
Lighting cycles synchronize (entrain) mammalian circadian rhythms by altering activity of cells in the suprachiasmatic nucleus (SCN) of the hypothalamus, a circadian pacemaker. Exposure of hamsters and rats to light pulses at those phases of the circadian rhythm during which light can shift the rhythm caused increased immunoreactivity for the product of the immediate-early gene c-fos in cells in the region of the SCN that receives retinal fibers. Light pulses also increased messenger RNA for the Fos protein and for the immediate-early protein NGFI-A in the rat SCN. Similar increases in mRNA for NGFI-A were seen in the SCN of hamsters. Thus cells in this portion of the SCN undergo alterations in gene expression in response to retinal illumination, but only at times in the circadian cycle when light is capable of influencing entrainment. 相似文献
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长末端重复序列(LTR)反转录转座子广泛存在于植物基因组中,本质是一段可移动的脱氧核糖核酸(DNA)序列。大多数LTR反转录转座子在外界环境变化下能够被激活转录,对环境变化做出响应。为研究毛竹基因组中的LTR反转录转座子的转录活性及在非生物环境胁迫下表达量的具体变化,克隆和鉴定了1个毛竹Phyllostachys edulis反转录转座子PHRE7。该转座子全长为6 073 bp,属于Ty1-copia家族中的Tork分支,LTR序列相似性为96.7%,插入时间为126.923万a前。对毛竹实生苗分别进行辐照(30,50,70 Gy),甲基化抑制剂(50,100,150 μmol·L-1),高温(42℃),低温(4℃),高盐(0.1,0.2,0.3 mol·L-1)等5种不同胁迫处理,通过定量荧光聚合酶链式反应(PCR)检测,PHRE7在INT,RT和RH等3个结构域中的表达量仅在辐照及0.2~0.3 mol·L-1高盐处理下随处理强度的上升而下降,其余所有处理(甲基化抑制剂、高温、低温、高盐0.1~0.2 mol·L-1)的表达量都随处理强度呈上升趋势。这些结果表明:PHRE7转座子是一个具有转录活性的LTR反转录转座子,且外界非生物环境胁迫对其表达模式有较大影响,表明PHRE7转座子能够响应外界环境变化。 相似文献
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Tissue-specific expression in transgenic mice of a fused gene containing RSV terminal sequences 总被引:25,自引:0,他引:25
Transgenic mice were generated with pRSV-CAT, a chimeric gene construct containing the long terminal repeat of Rous sarcoma virus (RSV) linked to the bacterial gene encoding chloramphenicol acetyltransferase (CAT). CAT expression, detected in adult animals of five independent strains, was preferentially directed to organs rich in tendon, bone, and muscle. This pattern reflects the disease specificity of the intact virus and suggests that the tissue tropism of RSV is determined at least in part by the presence of endogenous tissue-specific factors that can promote expression of genetic information linked to the long terminal repeat. In two of the mouse strains, insertion of the pRSV-CAT DNA resulted in developmental abnormalities. One of these strains was characterized by a dominant trait of embryonic lethality, the other by a recessive trait of fused toes in all four feet. 相似文献
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Molecular cloning of a feline leukemia virus that induces fatal immunodeficiency disease in cats 总被引:39,自引:0,他引:39
J Overbaugh P R Donahue S L Quackenbush E A Hoover J I Mullins 《Science (New York, N.Y.)》1988,239(4842):906-910
A replication-defective variant of feline leukemia virus was molecularly cloned directly from infected tissue and found to induce a rapid and fatal immunodeficiency syndrome in cats. Studies with cloned viruses also showed that subtle mutational changes would convert a minimally pathogenic virus into one that would induce an acute form of immunodeficiency. The data suggest that acutely pathogenic viruses may be selected against by current methods for isolation of the human and simian immunodeficiency viruses. 相似文献
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影响动物肠道SI基因表达的因素 总被引:1,自引:0,他引:1
Sucrase-isomaltase(SI,蔗糖酶异麦芽糖酶复合物,EC 3.2.1.48-10),是动物体内很重要的二糖酶,位于小肠刷状缘细胞膜上.SI合成的前体是分子量为260kD左右的肽链,加工成熟后同时具有蔗糖酶和异麦芽糖酶活性.SI对碳水化合物的肖化起重要作用.小肠肠道中SI含量受多种因素的影响,它们主要是通过调节SI基因的表达得以实现.本文综述了SI基因的特点以及碳水化合物、激素对其表达的影响.因SI基因是只在肠上皮细胞中表达的,所以对SI的研究有助于我们了解小肠酶的合成分化情况以及肠道特异基因表达的模式. 相似文献
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