High amplification of a densovirus-derived vector in larval and adult tissues of Drosophila.

The Lepidopteran densovirus-derived vector, pJlacZDeltaNS3, is a defective virus genome with an insertion of lacZ DNA in the viral structural protein coding sequence, and a deletion of the sequence coding the non-structural polypeptide NS3. pJlacZDeltaNS3 was injected into Drosophila eggs and the maintenance of the viral genome was monitored by expression of beta-galactosidase and by Southern blot hybridizations. Intense beta-galactosidase activity was observed in many somatic tissues of third-instar larvae and adult flies, in more than 60% of the injected animals. DNA analyses showed that staining in adult tissues correlated with the amplification of the vector. Together, these results suggest the occurrence of early events of integration of the vector into the Drosophila host genome.

Molybdoenzyme biosynthesis in Escherichia coli: in vitro activation of purified nitrate reductase from a chlB mutant.

All molybdoenzyme activities are absent in chlB mutants because of their inability to synthesize molybdopterin guanine dinucleotide, which together with molybdate constitutes the molybdenum cofactor in Escherichia coli. The chlB mutants are able to synthesize molybdopterin. We have previously shown that the inactive nitrate reductase present in a chlB mutant can be activated in a process requiring protein FA and a heat-stable low-molecular-weight substance. We show here that purified nitrate reductase from the soluble fraction of a chlB mutant can be partially activated in a process that requires protein FA, GTP, and an additional protein termed factor X. It appears that the molybdopterin present in the nitrate reductase of a chlB mutant is converted to molybdopterin guanine dinucleotide during activation. The activation is absolutely dependent upon both protein FA and factor X. Factor X activity is present in chlA, chlB, chlE, and chlG mutants.