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革兰氏阴性菌和革兰氏阳性菌的电泳迁移率:电动分析

The electrophoretic mobility of gram-negative and gram-positive bacteria: an electrokinetic analysis.

作者信息

Bayer M E, Sloyer J L

机构信息

Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA 19111.

出版信息

J Gen Microbiol. 1990 May;136(5):867-74. doi: 10.1099/00221287-136-5-867.

Abstract

The electrophoretic mobility (EPM) of a variety of Gram-negative and Gram-positive bacteria was measured with a Penkem S3000 analyser. Under standard growth conditions and neutral pH all cells displayed a negative EPM. The polysaccharide capsules of Escherichia coli strains K1, K5, K29 and K30 generated the highest EPM; to a lesser and varying degree O-antigens with charged groups and core lipopolysaccharides also contribute to the net EPM. Very little negative EPM was measured in suspension cultures of the gliding bacterium Cytophaga U67. No difference in the EPM was observed between rapidly growing and stationary-phase E. coli B. De-energization of the cell membranes by carbonyl cyanide m-chlorophenylhydrazone (CCCP) did not affect the EPM of wild-type and deep rough mutants of E. coli; and the EPM of Cytophaga U67 and Acholeplasma laidlawii remained unaltered by CCCP when measured in their respective growth media. Extrusion of filamentous bacteriophage f1 from cells of its host, E. coli A95, caused a shift to a higher negative EPM. We also measured a variety of Gram-positive strains, all of which displayed different EPMs. When membrane fractions of E. coli were adsorbed to latex spheres, characteristic differences between the EPM of beads coated with either inner or outer membrane were observed. The results suggest that the rapid EPM analysis is a useful tool to study the net electric charge of microorganisms and to examine changes of surface properties during interaction of cells with viruses, proteins (antibody) and charged antibiotics.

摘要

使用Penkem S3000分析仪测量了多种革兰氏阴性菌和革兰氏阳性菌的电泳迁移率(EPM)。在标准生长条件和中性pH下,所有细胞均显示负EPM。大肠杆菌K1、K5、K29和K30菌株的多糖荚膜产生了最高的EPM;带有带电基团的O抗原和核心脂多糖也在不同程度上对净EPM有贡献。在滑行细菌噬纤维菌U67的悬浮培养物中测得的负EPM非常少。在快速生长的大肠杆菌B和稳定期的大肠杆菌B之间未观察到EPM的差异。羰基氰化物间氯苯腙(CCCP)使细胞膜去能,这并不影响大肠杆菌野生型和深粗糙突变体的EPM;当在各自的生长培养基中测量时,CCCP对噬纤维菌U67和莱氏无胆甾原体的EPM没有影响。丝状噬菌体f1从其宿主大肠杆菌A95的细胞中挤出,导致EPM向更高的负值转变。我们还测量了多种革兰氏阳性菌株,它们均显示出不同的EPM。当大肠杆菌的膜组分吸附到乳胶球上时,观察到包被内膜或外膜的珠子在EPM上的特征差异。结果表明,快速EPM分析是研究微生物净电荷以及检查细胞与病毒、蛋白质(抗体)和带电荷抗生素相互作用期间表面性质变化的有用工具。

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