Four tandem defective P elements associated with positive regulation of theDrosophila melanogaster glucose-6-phosphate dehydrogenase gene

Three high-glucose-6-phosphate dehydrogenase (G6PD)-activity mutants (2512H, S44H, and 1FH) are characterized by two insertion sequences associated with the G6PD locus; one (Ins1; 3.5 kb long in 2512H and S44H and 2.9 kb long in 1FH) is present just 5' to exon I and consists of a KP' (the 32nd base of the KP was replaced by guanine), a core sequence and a KP, and the other is 4.2 kb long and resides within an intron. Southern blot analyses of revertants showing low G6PD activity suggested that the insertion sequence responsible for high G6PD activity may be the core sequence but not the flanking KP and KP' or the Ins2. DNA sequencing data of the clone carrying the core sequence of 2512H demonstrated that the core sequence is another type of defective P elements (core P). Interestingly, a protein(s) was found in the nuclear extract of Canton S embryos that specifically binds to the core P but not to the KP or various fragments of p pi 25.1. In addition, the mutant G6PD activity was found to be affected not only by the genotype, but also by cytoplasmic factors.     

Functional analyses of the maize CKS2 gene promoter in response to abiotic stresses and hormones

In our previous research, we showed that the cyclin-dependent kinase regulatory subunit (CKS2) in maize (Zea mays L.) was induced by water deficit and cold stress. To elucidate its expression patterns under adversity, we isolated and characterized its promoter (PZmCKS2). A series of PZmCKS2-deletion derivatives, P0–P3, from the translation start code (?1,455, ?999, ?367, and ?3 bp) was fused to the β-glucuronidase (GUS) reporter gene, and each deletion construct was analyzed by Agrobacterium-mediated steady transformation into Arabidopsis. Leaves were then subjected to dehydration, cold, abscisic acid (ABA), salicylic acid (SA), and methyl jasmonic acid (MeJA). Sequence analysis showed that several stress-related cis-acting elements (MBS, CE3, TGA element, and ABRE) were located within the promoter. Deletion analysis of the promoter, PZmCKS2, suggested that the ?999 bp promoter region was required for the highest basal expression of GUS, and the ?367 bp sequence was the minimal promoter for ZmCKS2 activation by low temperature, ABA, and MeJA. The cis-acting element ABRE was necessary for promoter activation by exogenous ABA.     

cis-Acting Sequences Required for Simian Foamy Virus Type 1 Vectors

We have constructed a series of vectors based on simian foamy virus type 1 (SFV-1) to define the minimum cis-acting elements required for gene transfer. To characterize these vectors, we inserted the coding sequence of the bacterial lacZ gene linked to the cytomegalovirus immediate-early gene promoter. Introduction of a deletion mutation in the leader region between the 5′ long terminal repeat and the start of the gag gene at position 1659 to 1694 completely abrogated gene transfer by the SFV-1 vector. Deletion of 39 nucleotides from position 1692 to 1731 in the leader region resulted in a significant reduction in the transducing-particle titer. Furthermore, we have identified a second cis-acting element located at the 3′ end of the pol gene between position 6486 and 6975 to be critical for SFV-1 vector transduction. These results identify the two important cis-acting elements required for SFV-1 vector construction, and the finding of a cis-acting element in the pol gene is unique among retroviruses.