Cloning And Expression Analysis Of Histone1(H1) Gene During Testis Development In Eriocheir Sinensis

Eriocheir sinensis(Crustacea: Decapoda, Grapsidae) is one of the important economic crabs in our country. In order to research the effect of H1 protein in uncondensation spermatozoa nuclear and the dynamic changes of H1 expression in spermatogenesis of Eriocheir sinensis, we obtained the c DNA sequence of H1 gene using PCR and RACE mathods. Through bioinformatics methods the gene was analyzed, which include amino acid analysis, tertiary structure analysis and comparison, homology analysis, phylogenetic tree analysis and genome structure analysis. At the same time, the expression of H1 and H1-delta gene in different testis development periods of Eriocheir sinensis was analyzed using real time RT-PCR.This protein plays an important role in establishing higher-order chromatin structure and regulating gene expression. So, it may be closely related to the morphology of mature sperm nuclear. In this study, an 804 bp full-length c DNA of Eriocheir sinensis was cloned, containing a 540 bp open reading frame(ORF) encoding a protein of 179 amino acids(18.279 k Da, p I 10.93), which displayed a reasonable degree of identity with the known H1 and shared the same canonical motif. This is the first study identifying, cloning, and sequencing the full-length c DNA of the H1 gene from E. sinensis. It showed that the structure of the H1 field was conservative by comparing the tertiary structure.The results of real-time RT-PCR revealed that the expression of H1 gene in spermatogonium stage, spermatocyte stage, spermatid stage of E. sinensis testis had significant differences compared with sperm period(P<0.05), but the expression of H1-delta only in spermatocyte stage had significant differences compared with sperm period(P<0.05), and the amount of expression all reached maximum at spermatocyte period and at sperm period dropped to the lowest. These results will provide the fundamental data for further explore of histone H1 dynamic change during E. sinensis spermatogenesis.