Zafarullah M, Wisniewski J, Shworak N W, Schieman S, Misra S, Gedamu L
Department of Biological Sciences, University of Calgary, Canada.
Eur J Biochem. 1992 Mar 1;204(2):893-900. doi: 10.1111/j.1432-1033.1992.tb16709.x.
A rainbow trout major heat-shock-protein-like gene (hsp 70) and corresponding cDNA clones were isolated by hybridization to heterologous hsp70 probes. DNA sequencing revealed that this gene is structurally similar to a mammalian heat-shock-cognate hsc70 gene and consists of eight introns. Northern blot and primer extension analyses showed that the corresponding mRNA is constitutively abundant in different trout tissues and salmonid cell lines. Fragments of the isolated gene containing the -900 - +30 and -217 - +58 sequence were linked to a bacterial chloramphenicol acetyltransferase reporter gene and transiently transfected into salmonid cells. The expression pattern of these constructs supports our conclusion that the isolated genomic and cDNA clones correspond to a trout heat-shock-cognate hsc70 gene.
通过与异源hsp70探针杂交，分离出了虹鳟鱼的一个主要热休克蛋白样基因（hsp 70）及相应的cDNA克隆。DNA测序表明，该基因在结构上与哺乳动物的热休克同源hsc70基因相似，由八个内含子组成。Northern印迹和引物延伸分析表明，相应的mRNA在不同的鳟鱼组织和鲑科细胞系中持续大量存在。将含有-900 - +30和-217 - +58序列的分离基因片段与细菌氯霉素乙酰转移酶报告基因相连，并瞬时转染到鲑科细胞中。这些构建体的表达模式支持了我们的结论，即分离出的基因组和cDNA克隆对应于一个鳟鱼热休克同源hsc70基因。
