Research On Six Genes Related To Egg-production In The Ovaries Of Zi Goose

In order to discuss the candidate genes associated with egg production and furtherly select marker-assisted, In the experiment we take a study of 6 genes associated with egg production. The ovary tissue of Zi-goose during prelaying 5-month and laying periods 8-month were taken as material in the experiment. Extracted the total RNA of ovary tissue two-month aged, acquired cDNA through reverse transcription and amplifed label aim fragments of the 6 genes expression succession by RT-PCR. First, measured the mRNA expression levels of the 6 unknown genes EST1～EST6 found in ovary tissue during prelaying and laying periods determined by the SYBR Green I Real-time quantitative PCR. Then, designed specific primers for the 5 genes EST2-EST6 and cloned respectively. Acquired the DNA fragments by sequencing. Application of software related with molecular biology to carry out bioinformatics analysis. In the end, through the RACE technology, we acquired the total cDNA sequence of a-enolase gene, predict and initially identify the function of the genes with molecular biology relative software. The results showed:1.The result of Real-time quantitative PCR showed that the expression level of EST1, EST2, EST4, EST5 and EST6 in the ovary of Zi-goose 8-month aged were higher than prelaying significatly (P<0.05); The relative expression of EST3 in laying period was 4.32 times prelaying period. The difference was extremely significant (P<0.01).2.The length of the egg production DNA fragments of the genes EST2-EST6 was 364bp,476bp,901bp, 908bp and 1 168bp. Compared the nucleotides homology and study the bioinformatic of the genes EST2-EST6, and acquired the basic material for cloning the total gene.3.In the experiment, we acquired the total cDNA sequence of a-enolase gene in Zi-goose's ovary tissue. The result of bioinformatic analysis showed the sequence contained a intact open reading frame (ORF), and the sequence length is 1 305bp, the 28 is start codon, the 1 332 is the terminate codon, there are 434 amino acids in the protein encoded by the gene, and there is anolase conserved domain in 5 to 416, located in cytoplasm, belong to cytoplasmic proteins. It contained a signal peptide cleavage site (between 20-21), include 2 inner helix across membrane and 1 outer helix across membrane. Establish the basement for identify the genetic function of a-enlose gene and furtherly studying the mechanism relate to egg production.