Creb Gene Cloning And Analysis And The Effect Of Melanin Synthesis Inhibitor And Reinforcer On Growth Traits Of The Larvae In Pteria Penguin

Pteria penguin is a large-type species with all black shells that can produce high quality of seewater pearl. Its expression level of melanin can affect the nacre color, thereby affecting the quality of pearls.In this study, the expression of melanin synthesis related genes Tyr, Creb, Cdk2, Pax3, Sox10, Bcl-2 in various tissues of Pteria penguin were analysed,Creb gene was cloned and sequenced in the first time, preliminarily studied the effect of melanin synthesis inhibitor and reinforce on growth traits of the larvae of Pteria penguin.Results are as follows:1.Studied the expression of melanin synthesis related genes Tyr, Creb, Cdk2, Pax3, Sox10, Bcl-2 in 7 tissues of Pteria penguin with Semi quantitative analysis,include mantle, gill, adductor muscle, digestive gland, testis, ovary and foot. The results showed: Tyr has the highest expression in mantle and testis,also expressed in Gill, adductor muscle and ovary,as it was related to the formation of nacre color. Bcl-2 highly expressed in mantle, adductor muscle, and testis, not expressed in digestive gland and ovary. Creb expressed in all tissues, but had higher expression in digestive gland, foot, testis and ovary. Cdk2 had higher expression level in digestive blind and testis,while low expressed in 5 other tissues. Pax3 only expressed in foot tissue, the expression of Sox10 was higher in the gills and digestive gland, low in testis and ovary.2. The Creb cDNA sequence of Pteria penguin was cloned for the first time with RACE-PCR technology.The total length of the Creb cDNA in Pteria penguin was 1479 bp, Open reading frame length was 1143 bp, encoding 356 amino acids. The Creb amino acid sequence contained a bZIP domain in 278~341 amino acid residues, had 4 protein binding sites, respectively in 1,253~254,273,356 amino acid residues, had 1 poly nucleotide binding site located in 296 amino acids. Homology analysis indicated that the deduced Creb amino acid sequence of Pteria penguin had 46%, 45%, 46% consensus with Crassostrea gigas, Crassostrea ariakensis, Ostrea edulis respectively,had 50%, 33%, 30% consensus with Lymnaea stagnalis,Aplysia californica,Biomphalaria glabrata,respectively.3. 1-month-old Pteria penguin larvae were treated with melanin synthesis inhibitor arbutin and reinforcer valproic acid using different concentration and treated time. After treated,cultured larvae in laboratory,observe the change of shell color, and analysis the effect of treatments to Pteria penguin shell length(SL), height(SH) and shell width(SW). The results showed that the larvae shell length、shell height、shell width growth of groups treated with different concentration and time of the arbutin were significantly higher than those of control groups,valproic acid treated groups were slightly higher than the control group; The larvae of treated group and control group in the first 90 days and 150 days increased rapidly in SL, SH, SW, but after that,from the 150 th day to the 200 th day, treatment groups and control groups SL, SH, SW grew slowly.The results indicated that arbutin and valproic acid had no inhibitory effect on the larvae of SL, SH, SW, but the treatment groups grew slightly faster than the control groups. The study showed that arbutin-treated groups had individuals with lighter shell color obviously, Valproic acid-treated groups had individuals with darker shell color. The result indicted that arbutin and valproic acid made the shell color of Pteria penguin larvae lighter and darker respectively in the growth process.Arbutin with the concentreations of 0.4 mM,2 mM, 4 mM and valproic acid with the concentreations of 1 mM, 5 mM and 10 mM all had no inhibition on SL, SH, SW of the larvae.