Growth Related Gene Cloning And Expression Of Apostichopus Japonicus

Sea cucumber(Apostichopus japonicus), commonly known as sea cucumber, belonging Echinodermata(Echinodermata), Holothuroidea(Holothuroidea), through hand mesh(Aspidochirota), Stichopodidae(Stichopodidae), imitation oplopanax(Apostichopus). Sea cucumber is a very important marine economy echinoderms, sea cucumber research of the past few years, the growth has made great progress, especially in water temperature, salinity, light, attached to the base, stocking density and food and other environmental factors on thorn Growth parameters related research and has more depth, in terms of genetic and developmental biology studies also have some progress. However, molecular genetics research on the growth area is very small.Cyclin B(Cyclin B), follistatin(Follistatin, Fst) and lower effector proteins(Tensilin) gene is related to the regulation of genes involved in growth, to a significant role in the growth and development of the sea cucumber. Cyclin B mainly achieved by controlling cell division G2 / M transition point to regulate the cell division, it is a eukaryotic cell cycle plays a key role in operation of regulatory factors. Follistatin is a rich glycosylated single polypeptide chain cysteines, is one of the transforming growth factor superfamily β, inhibit the secretion of follicle stimulating hormone(FSH) in. Under the effect of protein is a collagen fibronectin, collagen is produced by internal sea cucumber dermis layer of mutation.In this experiment, sea cucumber as raw material, the use of molecular biology techniques, the first cloned cyclin B, follistatin, tensilin genes obtained three cDNA Sequence. By bioinformatics analysis, cyclin B gene contains a 717 bp open reading frame encoding a 238 amino acid protein, follistatin gene contains a 1254 bp open reading frame encoding a 417 amino acid protein, tensilin gene It contains an open reading frame encoding a 273 amino acid protein consisting of a 822 bp.These three genes encoding proteins predictive analysis. Cyclin B includes a CYCLIN functional domains, without signal peptide transmembrane domain. Follistatin include six functional domains, according to the order are FOLN, KAZAL, FOLN, KAZAL, FOLN, KAZAL domain signal peptide transmembrane domain. Tensilin NTR includes a functional domain, transmembrane signal peptide domain.Real-time quantitative PCR(Real-Time qPCR) technology, cyclin B, follistatin, tensilin gene expression studies in different tissues of sea cucumber size, respectively. We found that the relative expression of cyclin B gene in an individual to be slightly higher than the sea cucumber in a small expression of the individual, body fluids highest expression of 1.103, 0.023 minimum body wall. follistatin gene in individual small sea cucumber in the relative expression levels of the organizations to be slightly higher, thebody wall up to 1.14, the lowest was 0.047 respiratory tree. tensilin gene expression of respiratory tree up to 1.14, the lowest expression of body fluid is 0.067.Use genome walking(Genome Walking) method, the cyclin B gene promoter cloning, has been the promoter sequence 2564 bp, the use of a promoter transcription binding site prediction programs are forecasted results showed a 7 binding sites, respectively, v-Myb, HNF-1, Evi-1, Elk-1, NF-Y, HNF, USF.