EXPRESSION OF PLACENTAL GROWTH HORMONE RELATED PROTEINS

休假培训的总体目标将是合成重组 大鼠胎盘生长激素/催乳素相关蛋白（GHRPs）， 表征它们对不同的 受体的形式。 我的实验室最近发现了一个新家族 GHRPs在大鼠胎盘中的作用，并由NIH资助研究GHRPs的作用 胎儿胎盘的生长 休假的目的是 GHRP家族中的单个蛋白质，评估它们各自的能力， 结合和激活不同形式的受体，并建立一个 预测妊娠大鼠潜在促生长作用的基础。 第一个具体目的是产生稳定表达以下的细胞系： 大鼠胎盘GHRPs和催乳素样蛋白B和 C. 将对含有重组蛋白的培养基进行评价， 与天然生长激素和催乳素受体的结合活性，如 以及已在Dr. 凯利的实验室。 休假的第一个学期将从9月开始 1992-1993年2月 我在佛罗里达大学的实验室 随后纯化由重组蛋白合成的重组蛋白， 在1992年3月至8月期间转染细胞系。 第二 具体目的是表征纯化的重组蛋白， 使用克隆的长和短形式的 在CHO细胞中表达的受体。 功能性将使用 不同PRL受体cDNA与β-乳珠蛋白的共转染 氯霉素乙酰转移酶（CAT）受体基因启动子 构建体 这项工作将在第二届任期内进行。 1993年9月至11月在法国休假。 我在佛罗里达的实验室 随后开始表征纯化的重组蛋白， 对孕鼠的促生长作用 1994. 在1994年9月至11月的第三个休假期内， 第三个具体目标是合成突变形式的重组 胎盘蛋白质使用定点诱变。 的结合和 突变蛋白的生物活性将被评估， 在凯利博士的实验室里克隆受体 突变的蛋白质 随后进行纯化并评价其在 佛罗里达大学的几个低出生体重的动物模型。

The overall goal of sabbatical training will be to synthesize recombinant rat placental growth hormone/prolactin related protein (GHRPs) and to characterize their binding and biological activities toward different forms of receptors. My laboratory has recently identified a novel family of GHRPs in rat placenta and is funded by NIH to study the role of GHRPs in feto-placental growth. The goal of the sabbatical is to synthesize individual proteins in the GHRP family, evaluate their respective ability to bind and activate different forms of receptors, and to establish a basis for predicting potential growth-promoting effects in pregnant rats. The first specific aim is to generate cell lines which stably express cDNAs for the rat placental GHRPs and the Prolactin-Like-Proteins B and C. Culture media containing recombinant proteins will be evaluated for binding activity to native growth hormone and prolactin receptors, as well as to cloned receptors which have been stably expressed in Dr. Kelly's laboratory. The first term of the sabbatical will be from Sep 1992-Feb 1993. My laboratory at the University of Florida will subsequently purify the recombinant proteins synthesized by the transfected cell lines during the period of Mar-Aug 1992. The second specific aim is to characterize the purified recombinant proteins for binding and biological activities using cloned long and short forms of receptors expressed in CHO cells. Functionality will be evaluated using cotransfection of different PRL receptor cDNAs with a Beta-lactoglobin promoter-chloramphenicol acetyltransfersase (CAT) receptor gene construct. This work will be carried out during the second term of the sabbatical in France from Sep-Nov 1993. My laboratory in Florida will subsequently begin to characterize the purified recombinant proteins for growth-promoting effects in pregnant rats during the period of Jan-Aug 1994. During the third term of the sabbatical, from Sep-Nov 1994, the third specific aim will be to synthesize mutated forms of recombinant placental proteins using site-directed mutagenesis. The binding and biological activities of the mutated proteins will be evaluated toward cloned receptors in Dr. Kelly's laboratory. The mutated proteins will subsequently be purified and evaluated for growth-promoting activity in several animal models of low birth weight at the University of Florida.