Expression Analysis Of Apostichopus Japonicus Elongator Protein2Gene During Regeneration Stages

Elongator which consists of ELP1-3core subcomplexes and ELP4-6smallersubcomplexes is a multiprotein complex. Elongator is highly conservative in yeast andhumans. It takes part in RNA polymerase II-mediated transcriptional elongation, tRNAmodification, cell division, cytoskeleton organization and plays an important role in theseprocesses. The elongator protein2(ELP2) is a member of the elongator multiprotein complex,therefore, it also participates in important physiological process in organisms, it is evryimportant and can’t be ignored in the growth and development of the organism.In this paper, the full-length cDNA of elongator protein2(Ajelp2) was cloned from thesea cucumber Apostichopus japonicus using rapid amplification of cDNA ends (RACE) PCRtechniques, it comprised3058bp, including a54bp5’ untranslated (UTR), a526bp3’ UTRand a2478bp open reading frame (ORF) encoding a polypeptide of825amino acids. Themolecular weight is91.6kDa and isoelectric of point presumptive protein is5.84. Throughthe Ajelp2gene sequence compares with other species, we find that it has high conservative.In situ hybridization test that where is the gene express in different organizations of the seacucumber Apostichopus japonicas, and the result show that the gene is expressed in eachorganization, especially it express level stronger in the body wall and intestines. In order todetect the gene expression level in the regeneration process of the sea cucumber Apostichopusjaponicas, we detect the expression patterns of the Ajelp2gene in different regenerationstages of different organization using real-time quantitative polymerase chain reaction(qRT-PCR). Experimental results show that the expression of the Ajelp2gene is similar in theintestine and respiratory tree, which is up-regulated with regeneration of organizations, on the15th day, the Ajelp2gene expression level is highest, then gradually down-regaled with theorganization returning to the formal size, and the Ajelp2gene expression level also returningto normal levels finally. In the body wall, the expression level of the Ajelp2gene was shortlyand obviously up-regulate in the early state and then down-regaled, but obvious up-regulatedat50th minute again. In the late state, the Ajelp2gene expression level gradually up-regulatewith regeneration of organizations and then return to normal levels.The experimental results show that Ajelp2gene acts as an important part during theregeneration of the sea cucumber Apostichopus japonicas. The study of the Ajelp2gene in thesea cucumber Apostichopus japonicas regeneration process provides reference material for the regeneration molecular mechanisms and regulation pathways in the future, and possesstheoretical significance, at the same time, our research have potential application value for theculturist understands the production, breeding, prevention, improve the survival rate of thelarval and other aspects.