Casey T A, Schneider R A, Dean-Nystrom E A
Physiopathology Research Unit, USDA-Agricultural Research Service, Ames, Iowa 50010.
Infect Immun. 1993 May;61(5):2249-52. doi: 10.1128/iai.61.5.2249-2252.1993.
A DNA probe derived from the subunit gene of a cloned 987P determinant was used to characterize the locations of the 987P genes in several Escherichia coli strains. We examined E. coli 987, a nonpiliated mutant of strain 987 (I36) that does not express 987P in vitro, and a variant of I36 that expressed 987P following growth in vivo. We found that plasmid and chromosomal copies of the 987P subunit gene could be differentiated in strain 987 by restriction endonuclease analysis and Southern blot hybridization. The nonpiliated mutant I36 had lost the plasmid copy but retained the chromosomal copy of the 987P gene. A 987P-piliated variant of I36, which did not contain the 987P plasmid, colonized and caused diarrhea in neonatal pigs similarly to wild-type 987. The plasmid that hybridized with the 987P probe was transferred from strain 987 to an E. coli K-12 strain by conjugation. We were unable to demonstrate expression of 987P by these transconjugants. The data suggest that the chromosomal and plasmid copies of the 987P genes may interact to influence 987P expression.
从克隆的987P决定簇的亚基基因衍生而来的DNA探针,被用于鉴定几种大肠杆菌菌株中987P基因的位置。我们检测了大肠杆菌987、987菌株(I36)的一个非菌毛突变体(该突变体在体外不表达987P)以及I36的一个变体(该变体在体内生长后表达987P)。我们发现,通过限制性内切酶分析和Southern印迹杂交,可以区分987菌株中987P亚基基因的质粒拷贝和染色体拷贝。非菌毛突变体I36丢失了987P基因的质粒拷贝,但保留了其染色体拷贝。I36的一个表达987P菌毛的变体,该变体不含有987P质粒,与野生型987一样,能在新生仔猪中定殖并引起腹泻。与987P探针杂交的质粒通过接合作用从987菌株转移到了一株大肠杆菌K-12菌株中。我们无法证明这些接合子能表达987P。数据表明,987P基因的染色体拷贝和质粒拷贝可能相互作用以影响987P的表达。