Expression Under Adversity And Cloning Analysis Of A Heat Shock Protein70 (HSP70) In Three Species Of Cultured Scallop

This study reported the variational expression of a heat shock protein70( HSP70)in three species of cultured scallop, Chlamys farreri, Argopectenirradians and Mizuhopecten yessoensis using immunological method. The sequencesof HSP70 cDNA from Argopecten irradians and Mizuhopecten yessoensis were alsodefined in this paper. By western immunoblotting,the effect of thermal stress on the expression ofHSP70 was obtained in various tissues such as hemolymph, muscle, gonad and gills.The results showed that HSP70 was prominently induced in gills, but not inhemolymph, muscle and gonad while exposed to heat shock. In the present study, flow cytometry (FCM) was for the first time introduced tomonitor HSP70 expression in hemolymph cells of scallops. In May 2003, hemolymphcells were collected from Chlamys farreri undergoing environmental stresses (heat,cold, bacteria infection and degradation).T-test showed that all those adverse factorsremarkably enhanced the production of HSP70(DF=8,P<0.01).Duncan-test showedthat: the blight of degradation environment (COD=8.36) approximated to injectionwith 20μlVibrio anguillarum(109cells/ml); heat aroused more notable increase ofHSP70 than cold under the same ?t. Same experiments were carried out in Oct.2003for all three species Chlamys farreri, Argopecten irradians and Mizuhopectenyessoensis. Duncan-test showed that heat aroused more remarkable increase of HSP70 3曲凌云 三种养殖扇贝热休克蛋白 HSP70 在逆境因子下的表达和相关基因的克隆 博士学位论文than cold under the same ?t in Argopecten irradians. But in Chlamys farreri andMizuhopecten yessoensis, the difference of effect was not distinct between those twostresses, heat and cold. Different results between spring and fall suggested that thethreshold induction temperature was changed with season in Chlamys farreri. FCMwas applied to track the induction of HSP70 in hemolymph cells of scallops after thetreat of heat shock. The analysis of these data led to the following observation: theHSP70 concentration in hemolymph cell was found to increase after heat stress;themaximum appeared at 7 hours later; and then decreased gradually. The HSP70 cDNAs were cloned by PCR,3'-RACE and 5'-RACE techniques from the gill tissues of Argopecten irradian and Mizuhopecten yessoensis(Partial Sequence Genbank Accession Number:AY435154,AY435155)。There is a coding sequence( CDS)of 2071 nucleotide acids coding 657amino acids in the sequence of HSP70 cDNA from Mizuhopecten yessoensis; and in the sequence of HSP70 cDNA from Argopecten irradians, the span of CDS is 2077 nucleotide acids coding 657amino acids. There is a 92.7% identity between HSP70 from Argopecten irradian and Mizuhopecten yessoensis in the amino acids sequence.