CLONING OF jnk1 AND jnk2 GENES AND EXPRESSION ANALYSIS IN RESPONSE TO HIGH TEMPERATURE STRESS IN LARIMICHTHYS POLYACTIS
JNKs(c-Jun amino-terminal kinases)has the function of regulating apoptosis and can respond to various extracellular signal stimuli.In order to study the regulatory mechanism of high temperature on liver cell apoptosis in Larimichthys polyactis,the apoptosis related gene jnk was investigated.The CDS sequences of jnk1 and jnk2 in Larimichthys crocea from NCBI were used as reference,the CDS sequences of these two genes in L.polyactis were cloned.Through bioinformatics analysis,it was found that the ORF(Open Reading frame)sequence of jnk1 in L.polyactis is 1 155 bp,encoding 384 amino acids,while the ORF sequence of jnk2 is 1 263 bp,encoding 420 amino acids.By predicting their structural domains,it was found that both jnk1 and jnk2 have a conserved STKc domain and a dual phosphorylation TPY catalytic site.Sequence alignment and phylogenetic analysis showed that both jnk1 and jnk2 in L.polyactis are closely related to those in L.crocea.By tertiary structure analysis,it is found that both jnk1 and jnk2 structures are conserved.Real-time quantitative PCR method was used to detect the distribution ofjnk1 and jnk2 in tissues,which showed that the expression levels of jnk1 and jnk2 in the brain of L.polyactis were the highest.Further analysis on the expression patterns of these two genes at different time under high temperature treatment revealed that there was no significant change in expression level for jnk1 with increasing treatment time.However,jnk2 exhibited significantly higher expression level than control group when the water temperature reached 32 ℃,but its expression gradually decreased with increasing treatment time,indicating an important regulatory role played by jnk2 during high temperature stress response process.These results provide a basis for exploring molecular mechanisms underlying liver response to high temperature stress and cell apoptosis regulation in L.polyactis.
Larimichthys polyactishigh temperature stressliverjnk1jnk2gene expression
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