Glutathione S-transferases(GSTs)are a superfamily of multifunction enzymes involved in the regulation of redox homeostasis and innate immune responses against various pathogenic infections in marine invertebrates.In t...Glutathione S-transferases(GSTs)are a superfamily of multifunction enzymes involved in the regulation of redox homeostasis and innate immune responses against various pathogenic infections in marine invertebrates.In the present study,a delta class GST gene(designated as FcδGST)was cloned from Fenneropenaeus chinensis using rapid amplification of c DNA ends(RACE)technology.The complete cDNA sequence of FcδGST was 780 bp in length,which includes a 27-bp 5′non-coding region(UTR),a 117-bp 3′UTR,a 636-bp open reading frame(ORF),and a polyadenylate signal site(AATAAA)presented at the upstream of poly A tail.The FcδGST gene encoded 211 amino acids peptide,including a GST_N domain and a GST_C domain,and exhibited high similarity with previously reported delta GSTs.The predicted molecular mass of FcδGST protein was 23.39 kDa,and its theoretical isoelectric point(pI)was 5.34.The FcδGST mRNA transcripts were ubiquitously expressed in all the tested tissues,with the highest expression level in hemocytes and hepatopancreas.During the stimulation of Vibrio anguillarum or white spot syndrome virus(WSSV),the m RNA expression of FcδGST in hemocytes and hepatopancreas revealed significant up-regulation.The purified recombinant FcδGST protein(designated as rFcδGST)exhibited specific catalytic activity against 1-chloro-2,4-dinitrobenzene(CDNB)substrate with relatively low stable enzymatic activities.These results indicated that FcδGST was a fragile but typical novel delta class GST member and potentially involved in the innate immune responses of F.chinensis.展开更多
基金Supported by the Project of Sanya Yazhou Bay Science and Technology City Management Foundation (No.SKJC-KJ-2019KY01)the Startup Fund of Young Talents Project of Ocean University of China。
文摘Glutathione S-transferases(GSTs)are a superfamily of multifunction enzymes involved in the regulation of redox homeostasis and innate immune responses against various pathogenic infections in marine invertebrates.In the present study,a delta class GST gene(designated as FcδGST)was cloned from Fenneropenaeus chinensis using rapid amplification of c DNA ends(RACE)technology.The complete cDNA sequence of FcδGST was 780 bp in length,which includes a 27-bp 5′non-coding region(UTR),a 117-bp 3′UTR,a 636-bp open reading frame(ORF),and a polyadenylate signal site(AATAAA)presented at the upstream of poly A tail.The FcδGST gene encoded 211 amino acids peptide,including a GST_N domain and a GST_C domain,and exhibited high similarity with previously reported delta GSTs.The predicted molecular mass of FcδGST protein was 23.39 kDa,and its theoretical isoelectric point(pI)was 5.34.The FcδGST mRNA transcripts were ubiquitously expressed in all the tested tissues,with the highest expression level in hemocytes and hepatopancreas.During the stimulation of Vibrio anguillarum or white spot syndrome virus(WSSV),the m RNA expression of FcδGST in hemocytes and hepatopancreas revealed significant up-regulation.The purified recombinant FcδGST protein(designated as rFcδGST)exhibited specific catalytic activity against 1-chloro-2,4-dinitrobenzene(CDNB)substrate with relatively low stable enzymatic activities.These results indicated that FcδGST was a fragile but typical novel delta class GST member and potentially involved in the innate immune responses of F.chinensis.
