Two remote and weak lac regressor binding sites can be used jointly to repress the synthesis of beta-galactosidase in E. coli, while they cannot separately. When this result is discussed in reference to the various modes of cooperation between the sites, it supports with a new approach a model implying the simultaneous binding of lac repressor to both sites with the formation of a DNA loop. In connection with this point, we present a new strategy to detect cooperative interactions in vivo, based on the asymmetry of the DNA binding site, formally equivalent here to a half-site, heterodimerization of the protein, and influence of orientation of the sites on repression at short and long distance.