Cloning and expression analysis of an inducible HSP70 gene from tilapia fish

Abstract

We isolated and characterized the tilapia (Oreochromis mossambicus) HSP70 gene, highly homologous to other HSP70 genes. A dramatic increase of tilapia HSP70 mRNA levels was observed after heat shock of whole animals in all organs tested. Reporter constructs were tested for transient expression in carp cells and in microinjected zebrafish embryos. The entire isolated regulatory region (3 3851/+157) was able to mediate heat shock inducible expression of the reporter gene, with no preference for a particular tissue. Our studies represent the first transcriptional analysis of a HSP70 promoter from fish, revealing a powerful tool to direct controlled, tissue-independent gene expression in fish.z 2000 Federation of European Biochemical Societies.

Gene structure and promoter function of a teleost ribosomal protein: a tilapia (Oreochromis mossambicus) L18 gene

“…We have also recently demonstrated tissue-independent expression of the L18 gene in O. niloticus by semiquantitative RT-PCR [18]. Consistent with these results, our microinjection experiments in zebra¢sh and tilapia embryos did not reveal any particular preference for a given tissue.…”

“…CAC36993) as a probe, 2U10 6 plaques (recombinant VGEM 11) of a tilapia (O. mossambicus) genomic library [17] were screened. Probe labeling, hybridization, phage DNA puri¢cation, subcloning and restriction mapping of positive clones were performed according to [18].…”

“…To obtain tiL18-0.5LacZ, this fragment was introduced into the expression vector pGCV-LacZ [20], where the SV40 promoter had previously been deleted [18]. The tiL18-0.2LacZ expression vector was obtained from the tiL18-0.5LacZ by digestion with NcoI and XhoI followed by religation.…”

<I>Mamestra brassicae</I> Nucleopolyhedrovirus Infection and Enhancing Effect of Proteins Derived From <I>Xestia c-nigrum granulovirus</I> in Larvae of <I>Mamestra brassicae</I> and <I>Helicoverpa armigera</I> (Lepidoptera: Noctuidae) on Cabbage

“…The HSFs bind to the HSEs and initiate the transcription of the HSP70s (Morimoto 1993). Although we did not measure the HSC70 or HSP70 protein level in this experiment, the precondition of higher levels of HSC70 or HSP70 is the higher abundance of HSC70 or HSP70 mRNA, respectively, because the synthesis of HSP70s is primarily regulated at the transcriptional level (Molina et al 2000). To sustain a proper level of HSP mRNA, the transcription of HSP genes can be controlled by negative regulation of HSPs.…”

Differential expression of three Paralichthys olivaceus Hsp40 genes in responses to virus infection and heat shock☆

“…So far, some Hsp genes have been cloned in different fish species, including Hsp70 from rainbow trout [25,26], medaka [27], zebrafish [28], tilapia [29] and pufferfish [30], Hsp90 from Chinook salmon [31] and Atlantic salmon [32], and several low molecular weight Hsps, including DnaJ homologues [25]. Heat shock is capable of inducing fish cells to increase the expression of Hsps [33].…”