Gene clones encoding phycocyanin and allophycocyanin were isolated from an Anabaena variabilis ATCC 29413-Charon 30 library by using the phycocyanin (cpc) genes of Agmenellum quadruplicatum and the allophycocyanin (apc) genes of Cyanophora paradoxa as heterologous probes. The A. variabiis cpcA and cpcB genes occur together in the genome, as do the apcA and apcB genes; the two sets of genes are not closely linked, however. The cpc and apc genes appear to be present in only one copy per genome. DNA-RNA hybridization analysis showed that expression of the cpc and apc genes is greatly decreased during nitrogen starvation; within 1 h no cpc or apc mRNA could be detected. The source of nitrogen for growth did not influence expression of the genes; vegetative cells from nitrogen-fixing and ammonia-grown cultures had approximately the same levels of cpc and apc mRNAs. Heterocysts had less than 5% as much cpc mRNA as vegetative cells from nitrogen-fixing cultures. Northern hybridization (RNA blot) analysis showed that the cpc genes are transcribed to give an abundant 1.4-kilobase (kb) RNA as well as two less prominent 3.8-and 2.6-kb species. The apc genes gave rise to two transcripts, a 1.4-kb predominant RNA and a minor 1.75-kb form.

When the filamentous, nitrogen-fixing cyanobacterium Anabaena variabilis ATCC 29413 was subjected to nitrogen starvation under aerobic conditions, a complex series of events was initiated which resulted in heterocyst formation and derepression of the ability to fix dinitrogen. Using DNA-RNA hybridization techniques, we monitored the expression of several genes during nitrogen starvation and correlated changes in the mRNA levels with changes in enzyme activity, protein levels, and morphology. Nitrogenase mRNA was first observed after about 8.5 h of nitrogen starvation, as was nitrogenase activity. Late proheterocysts were present at that time.