Expression of endogenous and microinjected hsp 30 genes in early Xenopus laevis embryos.
In the present study, we have examined the regulation of expression of a newly isolated member of the hsp 30 gene family, hsp 30C. Using RT-PCR, we found that this gene was first heat-inducible at the tailbud stage of development. We also examined the expression of two microinjected modified hsp 30C gene constructs in Xenopus embryos. One of the constructs had 404 bp of hsp 30C 5'-flanking region, whereas the other had 3.6 kb. Both gene constructs had 1 kb of 3'-flanking region. RT-PCR assays were employed to detect the expression of these microinjected genes. The presence of extensive 5'- and 3'-flanking regions of the hsp 30C gene did not confer proper developmental regulation, since heat-inducible expression of both of the microinjected constructs was detectable at the midblastula stage. The premature expression of the microinjected hsp 30 gene was not a result of high plasmid copy number of the presence of plasmid DNA sequences. These results suggest that the microinjected genes contain all the cis-acting DNA sequences required for correct heat-inducible regulation but do not contain the elements required for the proper regulation of hsp 30 gene expression during development. It is possible that regulatory elements controlling the developmental expression of the hsp30 genes may reside upstream or downstream of the entire cluster.
Ali
A
A
Department of Biology, University of Waterloo, Ontario, Canada.
Krone
P H
PH
Heikkila
J J
JJ
D000818:Animals; D004247:DNA / Q000235:genetics; D004277:DNA, Single-Stranded / Q000235:genetics; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005786:Gene Expression Regulation; D017343:Genes, Plant; D015698:Genomic Library; D006360:Heat-Shock Proteins / Q000235:genetics*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012045:Regulatory Sequences, Nucleic Acid; D017422:Sequence Analysis, DNA; D014162:Transfection; D014982:Xenopus laevis / Q000196:embryology* / Q000235:genetics*
D004277:DNA, Single-Stranded; D006360:Heat-Shock Proteins; D004247:DNA