MORPHOGENETIC PROCESSES IN BACTERIAL SPORULATION

形成独特的微生物结构所需的形态发生事件， B.subtilis孢子衣，将进行研究。特定的包膜多肽有 经过提纯，制备的抗体用作探针检测 前体、加工步骤和寻找被毛突变体中的变化。 二核苷酸残基交联包被多肽的机理 形成不溶于水的包衣比例有待进一步研究。抗体 将用于检测孢子壳结构蛋白基因的克隆。我们 还应尝试克隆影响萌发和孢子壳的基因 通过使用紧密相连的亮氨酸基因的克隆作为探针来构建。 在一个或多个指数后蛋白水解酶中改变的突变体将被分离， 绘制了图谱并构建了等基因菌株。这种物质的生理效应 蛋白质周转、适应下移和孢子被的突变 多肽(或其他蛋白质)的加工将进行研究。我们有 与后一事件有关的一种蛋白酶的证据，并应进行纯化 该酶测定的特异性、抑制性及体外研究 正在处理。我们还将进一步研究蛋白质的功能。 在孢子形成过程中早期产生的蛋白水解酶抑制物 使用改变后的抑制物的突变体。枯草杆菌谷氨酰胺合成酶 与许多其他生物不同的是，它似乎不受 复杂的级联机制也不受一系列调控基因的影响。全 到目前为止分离的突变体定位在结构基因内或接近结构基因，但 酶的产生量有一定的调节作用，尤其是由一种 在部分二倍体中可检测到的反式作用因子。通过适当的克隆 我们将定义这个反式作用因子，并对其进行排序 结构基因附近的潜在调控区。我们还将 构建更多的二倍体，与不同的类别进行互补研究 变种人。结构基因的克隆也将用于定位 突变体，尤其是构建缺乏任何 活性或抗原。这类突变体的多效性效应 将检查产孢量和分解代谢。

The morphogenetic events required to form a unique microbial structure, the B. subtilis spore coats, will be studied. Specific coat polypeptides are being purified, antibodies prepared and used as probes to detect precursors, processing steps and to look for alternations in coat mutants. The mechansim of cross linking of coat polypeptides via dityrosine residues to form an insoluble coat fraction will be further studied. The antibodies will be used to detect clones of spore coat structural protein genes. We shall also attempt to clone a gene affecting germination and spore coat structure by using as a probe a clone of the closely linked leucine genes. Mutants altered in one or more post exponential proteases will be isolated, mapped and isogenic strains constructed. The physiological effects of such mutations on protein turnover, shift down adaptation and spore coat polypeptide (or other protein) processing will be studied. We have evidence for one protease involved in the latter events and shall purify the enzyme to determine specificity, inhibition and to study in vitro processing. We shall also further examine the function of a protein protease inhibitor produced early during sporulation by trying to obtain mutants with an altered inhibitor. Glutamine synthetase in B. subtilis, in contrast to many other organisms, does not appear to be controlled by an elaborate cascade mechanism nor by a series of regulatory genes. All mutants isolated to date map in or close to the structural gene and yet there is some regulation of the amount of enzyme produced, especially by a trans acting factor detectable in partial diploids. By appropriate cloning we shall define this trans acting factor, and sequence it as well as potential regulatory regions close to the structural gene. We shall also construct more diploids to do complementation studies with various classes of mutants. Clones of the structural gene will also be used for localized nutagenesis, especially deletions to construct strains devoid of any activity or antigen. The pleiotropic effects of such mutants on sporulation and catabolism will be examined.