Characteristics Of Hira Gene And Its Promoter, And Its Annual Expression During The Development On Ovary In Crucian Carp, Carassius Auratus Var,Pingxiangnensis

Pingxiang red-transparent crucian carp,Carassius auratus var. Pingxiangnensis is a wildness natural triplod crucian carp mutant distributed in Pingxiang area of JiangXi Province. The fish is a good crucian carp species due to edible value with ornamental value, and is approved by national aquatic original improved variety committee as a new cultural fish species in2008. After successive generations of artificial selection, some good economic and genetic traits are basically stable to offspring, such as faster growth, more adaptability and higher nutritional value, easier to raise and breed, disease resistance and so on. The fish have two modes of reproduction,which includes bi-sexual and gynogenetic reproductive mode. Research show that the Hira gene is related to solution of the condensate about sperm nucleus in silver crucian carp, fruit flies and other organisms. This research as the material of the mutant fish, has a preliminary exploration on the Hira gene in fish reproductive role during the development. The major elements are as follow:1Using SMART cDNA synthesis and RACE-PCR techniques, the cloning of the full-length cDNA sequence of Hira was obtained, which contained an open reading frame of3033bp, encoding1011amino acids. The analysis of amino acid sequence showed that the Hira had a larger number of species specificity and stability.The Hira proteins of Pingxiang red crucian carp are highly homologous with the proteins of silver crucian carp, common carp and zebra fish, but distantly homologous to Amphibians, reptiles, birds and mammals. The analysis to Hira proteins showed that there are no signal peptide. Hira proteins may be soluble proteins and play role in the cytoplasmic matrix or nucleus. It seems to be in line with the characteristics of the regulation of gene transcription as histone chaperones.2We isolated a3.6kb fragment of the5'-flanking region (1.5kb) of Hira promoter from Pingxiang red-transparent crueian carp by Genome walking. The region contained one TATA box, one CAAT box and potential transcription factor binding sites including a EBOX, four OSP1and a POU1F1a. Both the sequence fragments and the core promoter region of the Hira, may be play a important role in the starting of the Hira gene transcription. 3The real-time quantitative polymerase chain reaction (qRT-PCR) was used to analyze the expression changes of the Hira gene of ovarian tissue during an annual developmenting. The results showed that the expression of Hira gene in ovarian tissue was prone to upward during an annual and was almost absent from the first month to the fifth. The relative amount of HIRA up to1947times at the6th month of age than the3th month, after sharply reduced. Then sharply decreased again and the expression of Hira reaching the peak at the11th month throughout the development of gonad. The relative amount of HIRA up to3383times than the3th month of age.