Institute of Molecular BioSciences, Massey University, Palmerston North, New Zealand.
Insect Mol Biol. 2012 Apr;21(2):169-80. doi: 10.1111/j.1365-2583.2011.01123.x.
In this study we report the isolation and characterization of a heat shock protein 70 (hsp70) gene, the hsp83 gene and two genes that encode small Hsps (Lchsp23 and Lchsp24) from the Australian sheep blowfly, Lucilia cuprina, a major agricultural pest. Phylogenetic analyses indicate that the LcHsp23 protein is the orthologue of Drosophila melanogaster Hsp23 and LcHsp24 is the orthologue of Sarcophaga crassipalpis Hsp23. Quantitative reverse-transcriptase PCR analysis showed that the basal level of Lchsp83 RNA is relatively high at all developmental stages and only moderately induced by heat shock. In contrast, Lchsp70 transcripts are present at low levels and strongly induced by heat shock at all stages. The basal levels of expression and degrees of heat induction of the Lchsp23 and Lchsp24 transcripts were more variable across the different developmental stages. Putative heat shock factor binding sites were identified in the Lchsp24, Lchsp70 and Lchsp83 gene promoters. The isolation of these hsp gene promoters will facilitate constitutive or conditional expression of a gene of interest in transgenic Lucilia.
在这项研究中,我们从澳大利亚羊狂蝇(Lucilia cuprina)中分离和鉴定了热休克蛋白 70(hsp70)基因、hsp83 基因和两个编码小热休克蛋白(Lchsp23 和 Lchsp24)的基因。澳大利亚羊狂蝇是一种主要的农业害虫。系统发育分析表明,LcHsp23 蛋白是黑腹果蝇 Hsp23 的直系同源物,LcHsp24 是 Sarcophaga crassipalpis Hsp23 的直系同源物。定量反转录聚合酶链反应分析表明,Lchsp83 RNA 的基础水平在所有发育阶段相对较高,仅适度地受到热休克的诱导。相比之下,Lchsp70 转录本的水平较低,但在所有阶段都强烈地受到热休克的诱导。Lchsp23 和 Lchsp24 转录本的基础表达水平和热诱导程度在不同的发育阶段变化较大。在 Lchsp24、Lchsp70 和 Lchsp83 基因启动子中鉴定出了推定的热休克因子结合位点。这些 hsp 基因启动子的分离将有助于在转基因 Lucilia 中组成型或条件性表达感兴趣的基因。