Reibach P H, Mask P L, Streeter J G
Can J Microbiol. 1981 May;27(5):491-5. doi: 10.1139/m81-072.
Bacteroids were isolated from the nodules of soybean plants by means of self-generating Percoll density gradients. The entire procedure can be performed in less than 1 h using an ordinary refrigerated centrifuge and angle head rotor. All of the markers for cytosol and bacteroid fractions behaved in accord with other reports in the literature. Asparaginase, beta-hydroxybutyrate dehydrogenase, and alanine dehydrogenase were all localized in the bacteroid fraction. Invertase, phosphoenolpyruvate carboxylase, and leghaemoglobin were all found in the cytosol fraction. Very little (less than 7%) cross contamination between the fractions was observed. The isolated bacteroids were viable, and based on electron micrographs, were free from contaminating plant material. Since the entire procedure is performed isosmotically, very little damage to the bacteroids is likely to occur. No organic compounds, except Percoll, were added to the isolating media, thus aiding in the analysis of bacteroid and cytosol metabolites.
通过自生成的Percoll密度梯度从大豆植株的根瘤中分离出类菌体。使用普通的冷藏离心机和角头转子，整个过程可在不到1小时内完成。胞质溶胶和类菌体组分的所有标志物的表现与文献中的其他报道一致。天冬酰胺酶、β-羟基丁酸脱氢酶和丙氨酸脱氢酶均定位于类菌体组分中。转化酶、磷酸烯醇丙酮酸羧化酶和豆血红蛋白均在胞质溶胶组分中发现。各组分之间的交叉污染非常少（不到7%）。分离出的类菌体是有活力的，根据电子显微镜照片，没有受到植物材料的污染。由于整个过程是在等渗条件下进行的，因此类菌体很可能很少受到损伤。除了Percoll外，分离培养基中未添加任何有机化合物，从而有助于分析类菌体和胞质溶胶代谢物。
