The Molecular Characteristics And Expression Analysis In Different Tissues And Different Ages Of ?-catenin From Hyriopsis Schlegelii

Hyriopsis schlegelii is a kind of high quality freshwater breeding mussel.The studyon its multiplication can provide a lot of theoretical basis for breeding.The study can also solve the problems of breeding at this stage.In order to explore the mechanism of sexual expression and controlling of freshwater shellfish,we established a gonadal transcriptome of Hyriopsis schlegelii.In the transcriptome library,the ?-catenin gene sequence was present.The ?-catenin gene was verified in this study.The full-length c DNA was cloned by RACE PCR,which named Hs?-catenin.The Hs?-catenin gene sequence and its protein were analyzed by different biological softwares.The Hs?-catenin c DNA presented as a full length of 4386 bp that contained an open reading frame(ORF)of 2466 bp coding a protein of 821 amino acids.The molecular weight of the protein was366046.5Da.The number of atoms was 47068.The molecular expression was C13499H22614N4386O5661S908.The protein was acidic.The most abundant amino acids were leucine(Leu),alanine(Ala)and serine(Ser),whose contents were 10.7%,8.8%and 7.5% respectively.The least amino acids were cysteine (Cys)and tryptophan(Trp),whose contents were 1.3% and 0.8% respectively.The content of basic amino acid was 11.5% and the content of acidic amino acid was 10.7%.The protein was mainly composed of 12 ARM repeat sequences.The ARM domain was the central conserved region of ?-catenin in each species,so that the accuracy of the experimental results was verified.In the secondary structure,the ?-helix who was distributed in large numbers accounted for 47.75%.The ?-folding accounted for1.22%.The random curl accounted for 51.04%.Tertiary structure had many ?-helixs which were also right-handed superhelixes.Every three ?-helixs composed a ARM domain.This protein had 12 ARM domains.Hs?-catenin gene was clustered with homologous genes from soft animal in the phylogenetic tree,including Pinctada martensii?Azumapecten farreri and Crassostrea gigas.Hs?-catenin was also highly homologous to insects,such as Chaetopterus variopedatus ?Platynereis dumerilii and Danaus plexippus.The homology with human and fish was low.The expression of Hs?-catenin gene was detected by real-time quantitative PCR(qRT-PCR),and the different expression of different age and different sex in tissuees was detected.The results showed that Hs?-catenin was expressed in all tissues.The expression from high to low were intestines,foot,testis,liver,ovary,heart,mantle,kidney and blood.This suggested that Hs?-catenin had a variety of biological functions in different tissues.It was higher in mature testis than that in mature ovary.We suggested that Hs?-catenin was more involved in other biological functions after gonad maturation.Gonad was appearance at 5 month.Male and female can be identified at 12 months.Gonad was matured at 36 month.The expression of Hs?-catenin in different ages and different gonads was detected.The results showed that the expression was highest in three-year-old(about 36 months).The expression of one-year-old(about 12 months)was following.The expression in tow-year-old(about 24 months)and four-year-old(about 48 months)was lowest.The expression level of ovary from one-year-old to three-year-old was significantly higher than that in testis.However the testis was slightly higher than ovary at four-year-old.These results indicated that it maight be involved in the regulation and differentiation of Pigeon mussel.The results of qRT-PCR showed that the highest expression of Hs?-catenin was in intestine.The expression level was high at the age of 12-month-old(gender differentiation)and 36-month-old(gonad mature),indicated that it might be involved in sex determination and sex differentiation of Hyriopsis schlegelii.