Molecular Cloning,Characterization And Tissue Expression Analysis Of The Ovine DNAJC28 Gene From Small-tail Han Sheep

Heat shock proteins(heat shock proteins,HSPs)are a group of ubiquitous in prokaryotes and eukaryotes,under stress protein expression was significantly increased.According to molecular size,heat shock protein can be divided into six subfamilies: Hsp100,Hsp90,Hsp70,Hsp60,Hsp40,and small heat shock proteins(eg HSP27).Wherein the Hsp40 With J domain is also known as DNAJ.In this study,we cloned and identified a new member of the Hsp40 family,DNAJ(Hsp40)homolog,subfamilyC,member28,called DNAJC28.Heat shock proteins are an important part of animal anti-stress response mechanisms,the process for sheep anti-stress plays a key role.DNAJ originally found in Escherichia coli,known as Hsp40 in eukaryotes,Hsp70 is a subclass cofactors,a major regulator of Hsp70 activity.DNAJC28 DNAJ gene is a gene family,for regulating the biological function of the corresponding protein Hsp70 plays a key role.In this study,RT-PCR,RACE,fluorescence quantitative methods such as cloning the cDNA sequence DNAJC28 gene and mRNA abundance and protein levels were analyzed.The main results are as follows:(1)In this study,RACE methods to Han sheep as experimental material,the full-length cDNA clone Han sheep DNAJC28 gene.The full-length cDNA is 2853 bp,an open reading frame encoding 380 amino acids.(2)Detected DNAJC28 genes showing differential expression in tissue Han sheep and Dorper sheep.Organizations in different tissues of expression differ greatly,detected in longissimus muscle biceps arm,intercostal muscle in the expression and the expression of a relatively small amount in the internal organs and other tissues.(3)DNAJC28 amino acid homology phylogenetic analysis revealed that the protein is highly conserved in mammals,the gene of sheep and cattle,goats similar.In summary,this study successfully cloned gene DNAJC28 Han sheep,the structure of the amino acid sequence encoded by the gene sequence and predicted,and comparative analysis of small-tail sheep and Dorper sheep DNAJC28 gene mRNA and protein the expression level of organization on the spectral characteristics for subsequent studies of the gene provides a certain basis.