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

研究者建议将分子生物学技术应用于 在LCMV诱导的抑制的基础上， 在持续感染的细胞中分化功能。 两个领域 建议中概述了检查。 第一个问题是 观察到的生长激素产生的抑制是否 持续感染动物的垂体是由于 生长激素mRNA的稳定状态水平或由于生长激素水平的降低， 生长激素mRNA的翻译。 第二个调查领域是 对持续感染的病毒RNA进行分析， 组织和细胞，以了解病毒的机制， 持久性。 这两种方法都将利用以下的北方印迹杂交： 从受感染的组织或细胞中提取的RNA。 这些污点 用从重组质粒制备的切口平移探针进行询问 含有部分大鼠生长激素cDNA的探针或用由 克隆的病毒核酸的L和S片段。 此外，GH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.