THE EFFECT OF LCM VIRUS INFECTION ON THE GENETIC REGULATION OF GH GENES

这位研究人员建议将分子生物学技术应用于 大鼠巨噬细胞病毒诱导抑制的基础研究 持续感染细胞的分化功能。以下两个领域 提案中概述了考试情况。第一个解决了这个问题 是否观察到生长激素分泌的抑制 持续感染的动物的脑下垂体是由于 生长激素mRNA的稳定水平是由于 生长激素基因的翻译。第二个调查领域是 对持续感染的人体内存在的病毒RNA进行分析 组织和细胞，以了解病毒的机制 坚持不懈。这两种方法都将利用Northern杂交技术 从受感染的组织或细胞中提取的RNA。污点将会是 用从重组质粒制备的尼克翻译的探针进行询问 含有部分大鼠生长激素的cDNAi或与由 克隆了L和S的病毒核酸片段。此外，生长激素合成 将通过分析多聚体mrna以及体外研究 感染细胞Poly(A)+RNA的翻译。

The investigator proposes to apply the techniques of molecular biology to the investigation on the basis of the LCMV-induced repression of differentiated function in persistently infected cells. Two areas of examination are outlined in the proposal. The first addresses the question of whether the observed depression in growth hormone production in the pituitaries of persistently infected animals is a result of an alteration in the steady state level of growth hormone mRNA or is due to reduction in the translation of growth hormone mRNA. The second area of inquiry is to conduct an analysis of the viral RNA present in persistently infected tissues and cells in order to understand the mechanisms of viral persistence. Both approaches will utilize Northern blot hybridization of RNAs extracted from infected tissues or cells. The blots will be interrogated with nick translated probes prepared from a recombinant plasmid containing a partial rat growth hormone cDNA or with probes prepared from cloned L and S segments of viral nucleic acid. In addition, GH synthesis will be studied by analysis of polysomal mRNAs as well as in vitro translation of the infected cell poly (A)+ RNA.