Total polysomal RNA and polyadenylated mRNA from third instar larvae, pupae, and adults of D. melanogaster were hybridized in vast excess to labeled single-copy DNA in order to measure the sequence complexity of each RNA population. Then, to measure the sequence homology between the populations, each was hybridized to DNA enriched for messenger coding sequences in third instar larvae and to DNA depleted of these sequences. Our results show that a similar number of genes, approximately 16,000, is expressed in larvae, pupae, and adults, and that only one-third of these is expressed as polyadenylated mRNA. Further, the composition of both polyadenylated and nonpolyadenylated mRNA classes is shown to change very little between these three stages of development. Finally, the head of adult Drosophila is shown to contain 11,000 RNA species, approximately 70% of the number contained in the entire adult. © 1981.

The expression of 20 cloned DNA sequences, selected on the basis of the prevalence of their transcripts in Drosophila adult head, was examined during Drosophila development. The amount of polyadenylated RNA complementary to these clones was measured in the cytoplasm of 0- to 20-hr embryos and on the polysomes of third instar larvae and 5- to 7-hr pupae. Four patterns of developmental expression were observed among the 20 clones, including 6 clones whose transcripts are prevalent only on polysomes of the adult head. Multiple genes closely clustered within the DNA of individual clones were shown to be expressed coordinately during development in some cases and differentially in others. © 1982.

Comparative screening of a library of cloned Drosophila DNA with polyadenylated mRNA from Drosophila adult head and adult body identified 20 cloned sequences expressed more abundantly in the tissues of the head than in other tissues. Quantitation using a "dot blot" hybridization assay demonstrated that the DNA sequences are expressed on adult head polysomes from 3 to 177 times more abundantly than on body polysomes, and their transcripts represent from 0.05 to 0.66% of the polyadenylated mRNA mass of the head. The steady-state nuclear RNA concentrations of four species were determined to be from 13 to 48 times greater in head nuclei than in body nuclei, an indication that their expression is controlled at least in part at the transcriptional level. The chromosomal locations of all 20 head-specific clones were identified by in situ mapping, and no distinct clustering was observed. However, four of the clones were shown by Southern and Northern blot analysis to contain multiple RNA coding sequences. The genes in these tightly clustered sets were observed to be expressed simultaneously in some cases and differently in others. © 1982.

The content and sequence complexity of the nuclear DNA and messenger RNA for epimastigotes of Trypanosoma cruzi were determined. From analysis of nuclear DNA reassociation studies and microspectrofluorometric measurements of laser induced fluorescence of cellular DNA, T. cruzi is found to be a diploid organism with a nuclear DNA content of 2.5 x 10(8) nucleotide pairs (2.8 x 10(-13) g) and a kinetoplast DNA content of 4.9 x 10(7) nucleotide pairs (5.4 x 10(-14) g). Reassociation kinetics of nuclear DNA of average length 0.4 kb reveals three kinetic components: a moderately repetitive component with a reiteration frequency of 5.1 x 10(3) present in 9% of the fragments, a lowly repetitive component with a reiteration frequency of 32 present in 51% of the fragments, and a single-copy component present in 23% of the fragments. By saturation hybridization of total polysomal RNA to 3H-labeled single-copy DNA, it was determined that 68% of the single-copy DNA was represented in the epimastigote polysomal RNA. This corresponds to ca. 12 000 different mRNA species. Of these, ca. 9000 are present as poly(A)+-RNA, while the remaining 3000 appear not to be polyadenylated. Kinetic analysis of the poly(A)+-RNA population indicates it is composed of at least three classes of RNA's of different abundancy levels: two sequences which occur ca. 3000 per cell, ca. 750 sequences which occur about 20 times per cell, and ca. 15 500 sequences which occur 1-2 times per cell.