Promoter analysis of the membrane protein gp64 gene of the cellular slime mold Polysphondylium pallidum.
We cloned a genomic fragment of the membrane protein gp64 gene of the cellular slime mold Polysphondylium pallidum by inverse PCR. Primer extension analysis identified a major transcription start site 65 bp upstream of the translation start codon. The promoter region of the gp64 gene contains sequences homologous to a TATA box at position -47 to -37 and to an initiator (Inr, PyPyCAPyPyPyPy) at position -3 to +5 from the transcription start site. Successively truncated segments of the promoter were tested for their ability to drive expression of the beta-galactosidase reporter gene in transformed cells; also the difference in activity between growth conditions was compared. The results indicated that there are two positive vegetative regulatory elements extending between -187 and -62 bp from the transcription start site of the gp64 promoter; also their activity was two to three times higher in the cells grown with bacteria in shaken suspension than in the cells grown in an axenic medium.
Takaoka
N
N
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo, Japan.
Fukuzawa
M
M
Saito
T
T
Sakaitani
T
T
Ochiai
H
H
D000818:Animals; D001483:Base Sequence; D015815:Cell Adhesion Molecules / Q000235:genetics*; D056890:Eukaryota / Q000235:genetics*; D017125:Genes, Protozoan; D008562:Membrane Glycoproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D015800:Protozoan Proteins / Q000235:genetics; D017422:Sequence Analysis, DNA; D014764:Viral Proteins
D015815:Cell Adhesion Molecules; D008562:Membrane Glycoproteins; D015800:Protozoan Proteins; D014764:Viral Proteins