摘要
A cDNA encoding heat shock cognate protein 70（HSC70）was cloned from liver of grass carp（Ctenopharyngodon idella）（GenBank JF436930）.This cDNA was found out to contain2 346 bp in length,including 1 950 bp of complete coding sequence encoding 649 amino acids（aa）,plus 89 bp of 5′-UTR and 307 bp of3′-UTR.Analysis of its genomic structure revealed that its corresponding gene contained seven exons and six introns.Homology analysis indicated that it shared 99%of identity with HSC70 of breams and 86%of identity with HSP70 of Drosophila.Fluorescent RT-PCR analysis revealed that at 28℃,this gene was expressed in abdominal fat,muscle,intestines,brain,middle kidney,head kidney,gonads,swim bladder,liver,heart,spleen,gills,and fins with expression level in liver being the highest（p〈0.05）,followed by that in the gonads;at 36℃,its mRNA expression level was increased at first but then decreased thereafter under heat shock stress,indicating that its expression can be regulated by heat shock.In conclusion,cloning and expression analysis identified a cDNA encoding a constitutive HSP70 gene that is expressed in many tissues of Ctenopharyngodon idella and its expression was down-regulated by heat shock.
A cDNA encoding heat shock cognate protein 70（HSC70）was cloned from liver of grass carp（Ctenopharyngodon idella）（GenBank JF436930）.This cDNA was found out to contain2 346 bp in length,including 1 950 bp of complete coding sequence encoding 649 amino acids（aa）,plus 89 bp of 5′-UTR and 307 bp of3′-UTR.Analysis of its genomic structure revealed that its corresponding gene contained seven exons and six introns.Homology analysis indicated that it shared 99%of identity with HSC70 of breams and 86%of identity with HSP70 of Drosophila.Fluorescent RT-PCR analysis revealed that at 28℃,this gene was expressed in abdominal fat,muscle,intestines,brain,middle kidney,head kidney,gonads,swim bladder,liver,heart,spleen,gills,and fins with expression level in liver being the highest（p〈0.05）,followed by that in the gonads;at 36℃,its mRNA expression level was increased at first but then decreased thereafter under heat shock stress,indicating that its expression can be regulated by heat shock.In conclusion,cloning and expression analysis identified a cDNA encoding a constitutive HSP70 gene that is expressed in many tissues of Ctenopharyngodon idella and its expression was down-regulated by heat shock.
基金
Supported by the Outstanding Young Scientist Research Award Fund of Shandong Province of China(BS2013NY002)
