Expression Analysis Of Apostichopus Japonicus Fibrinogen-like Protein A During Regeneration Stages

Fibrinogen-like protein A, a member of the fibrinogen-related protein superfamily, existsin different tissues of vertebrates and invertebrates. It plays important part in manyphysiological and biochemical reactions including innate immune response, wound healing,blood clotting, regeneration and so on.In this paper, RACE technology was performed to obtain the full-length cDNA sequenceof the Aj-fglA gene for the first time. The putative amino acids of this gene was analysed bybioinformatic. In situ hybridization was used to research different expression pattern ofAj-fglA in different tissues of sea cucumber. Real-time PCR was used to study the expressionlevel of Aj-fglA during different regeneration stages in the body wall, intestine and respiratorytrees.The clone analysis of FGLA showed: the full-length cDNA of Aj-fglA is composed of1524bp included a849bp opening reading frame (ORF) encoding a theoretic282amino acidprotein. And the bioinformation analysis showed that FGLA has a N-terminal signal peptideand a C-terminal FBG domain, a62bp5`untranslated region (UTR) and a613bp untranslatedregion in the3`UTR. The calculated molecular mass of the whole protein is31.96kDa and itspredicted isoelectric point is5.64. According to the characteristics of this protein, FGLA is amember of fibrinogen-related protein superfamily. In situ hybridization demonstrated that theexpression of Aj-fglA was located in the body wall, intestinum, longitudinal muscles andrespiratory tree. Real-time PCR data suggested that there was a remarkable up-regulation ofAj-fglA gene expression in the the early stage of body wall, the summit value appeared at10min and40min, and subsequent down-regulation. At the second stage of body wall, as thetissues regenerate, Aj-fglA gene expression level kept on increasing, the mix value appeared at5d, then came to the noamal level. In intestinum and respiratory tree the expression level ofAj-fglA was up-regulation as well, at20d and25d, a high level expression appearedrespectively. The expression level of Aj-fglA at all these regeneration stages were higher thanthat of normal tissues in four tissues, until the new organizations recovered, the expressionreached to normal. Thus it can be seen that Aj-fglA gene play a key role in regenerationprocess of Apostichopus japonicas.Sea cucumber (Apostichopus japonicas) was deeply loved by the people for its highnutrition value and commercial value. How to improve the growth rate and reduce the impact of vomiting, as well as improve the survival rate of larval are the main issues for maricultureof sea cucumber. Therefore, research the process of regeneration, regulatory pathways andrelevant molecular mechanism possess important theory significance and practical value.