Repeated immunization with plasmid DNA formulated in poly(lactide-co-glycolide) microparticles is well tolerated and stimulates durable T cell responses to the tumor-associated antigen cytochrome P450 1B1.

Injection of microparticle-encapsulated DNA elicits immune responses to plasmid-encoded antigens in mice and humans. Cytochrome P450 CYP1B1 (CYP1B1) is a member of the CYP1 P450 enzyme family that is overexpressed in a variety of solid tumors. The work described herein was performed to study the kinetics of stimulating T cell responsiveness with an encapsulated DNA encoding CYP1B1 and provides support for the clinical development of this formulation.

Mutation detection using nucleotide analogs that alter electrophoretic mobility.

A simple primer extension assay has been developed to distinguish homologous DNA segments differing by as little as a single nucleotide. DNA strands are synthesized with one of the four natural nucleotides replaced with an analog that affects electrophoretic mobility. DNAs that are the same length but differ in the number of analog molecules per strand exhibit different mobilities on a sequencing gel.

Proximity-dependent enhancement of Sgs-4 gene expression in D. melanogaster.

A weakly expressed allele of Sgs-4 obtained from the Kochi strain of D. melanogaster (Sgs-4K) increases 4-fold in levels of accumulated transcript when paired with a wild-type (Oregon-R) Sgs-4 allele (Sgs-4ORE) and increases 9-fold when paired with a duplication of the wild-type Sgs-4 allele Confluens (Sgs-4Co). There is no enhancement of expression when Sgs-4K is paired with the Sgs-4 null allele Ber-I (Sgs-4BER) or with another weakly expressed Sgs-4 allele, Hikone-R (Sgs-4HIK); there is minimal enhancement when chromosome pairing is disrupted near the Sgs-4 locus by rearrangement of the wild-type Sgs-4 allele.

Variegated expression of the Sgs-4 locus in Drosophila melanogaster.

Experiments on T(1;4)wm258-21 larvae of Drosophila melanogaster are described which establish the existence of a salivary gland specific marker for position-effect variegation. The marker is a glue protein gene called Sgs-4 which is expressed during the third larval instar. Using temperature as a variegation modifier, we showed that cytological compaction for the Sgs-4 chromosomal locus is enhanced at 17 degrees C and reduced at 29 degrees C.