Krone P H, Heikkila J J
Department of Biology, University of Waterloo, Ontario, Canada.
Development. 1989 Jun;106(2):271-81. doi: 10.1242/dev.106.2.271.
The expression of microinjected chimeric genes containing Drosophila hsp 70 and Xenopus hsp 70 and hsp 30 promoters linked to the reporter gene coding for bacterial chloramphenicol acetyltransferase (CAT) was examined during early development of Xenopus laevis. Heat-inducible expression of fusion genes containing either the Drosophila hsp 70 promoter (1100 bp) or the Xenopus hsp 70 promoter (750 bp) was first detectable after the midblastula stage of development. This coincides with the embryonic stage at which the endogenous hsp 70 gene is first heat-inducible. A Xenopus hsp 30/CAT fusion gene containing 350 bp of promoter sequences was also heat-inducible after the midblastula stage unlike the endogenous hsp 30 genes which were not heat-inducible until the early tailbud stage (stage 23-24). Sequences that are present within either the coding or 3' region of the hsp 30 clone do not cause the microinjected hsp 30 gene to be developmentally regulated in a normal manner. Additionally, microinjected hsp 30 gene sequences have no effect on the developmental regulation of endogenous hsp 30 genes which continue to be activated at the tailbud stage of development. Our data suggest, that an inhibitory system, which may control the expression of the endogenous hsp 30 gene during development, does not regulate the expression of the injected hsp 30 gene.
在非洲爪蟾早期发育过程中,研究了显微注射的嵌合基因的表达情况,这些嵌合基因包含果蝇热休克蛋白70(Drosophila hsp 70)、非洲爪蟾热休克蛋白70(Xenopus hsp 70)和热休克蛋白30(hsp 30)启动子,并与编码细菌氯霉素乙酰转移酶(CAT)的报告基因相连。含有果蝇热休克蛋白70启动子(1100 bp)或非洲爪蟾热休克蛋白70启动子(750 bp)的融合基因的热诱导表达,在发育的中囊胚期之后首次可检测到。这与内源性热休克蛋白70基因首次具有热诱导性的胚胎阶段相吻合。与内源性热休克蛋白30基因不同,含有350 bp启动子序列的非洲爪蟾热休克蛋白30/CAT融合基因在中囊胚期之后也具有热诱导性,内源性热休克蛋白30基因直到早期尾芽期(第23 - 24阶段)才具有热诱导性。热休克蛋白30克隆的编码区或3'区域内存在的序列,不会使显微注射的热休克蛋白30基因以正常方式受到发育调控。此外,显微注射的热休克蛋白30基因序列对内源性热休克蛋白30基因的发育调控没有影响,内源性热休克蛋白30基因在发育的尾芽期继续被激活。我们的数据表明,在发育过程中可能控制内源性热休克蛋白30基因表达的抑制系统,并不调控注射的热休克蛋白30基因的表达。