Study On The New Genes Of Sow Fertility

After corresponding analysis of the relationship with sow gestation,three Expressed sequence tag(CK455976,CK465614 and CO986776)on the top of Sow gestation relevant gene regulatory networks which was constructed by former researcher in our laboratory was selected to do forward research.Using the total RNA extracted from sow ovaries(85-days-pregnacy) and primers designed according to the sequence of these EST,cDNA complete sequence of these three genes have been obtained using RT-PCR and RACE-PCR technology.And after bioinformatics analysis, tissue expressed analysis and chromosome location,we got the results:(1) The full-length cDNA sequence from the porcine CK455976 were cloned,and the cDNA was 596bp with an entire open reading frame,encoding 117 amino acids.The sequences have been deposited in GenBank database,the accession number is FJ436413. The predicted nucleotide sequence similarity with human,mouse,dog and macaca mulatta DPPA5 gene were respectively 83%,70%,80%,83%,and the amino acid sequence were respectively 74%,55%,69%,74%.The INRA-Minnesota porcine radiation hybrid panel(IMpRH) were employed to determine the chromosomal locations of the DPPA5.The DPPA5 was localized to 1p1.3,with close linkage to SW1123 (LOD=5.82).(2) The full-length cDNA sequence from the porcine CK465614 was cloned.The sequences have been deposited in GenBank database,the accession number is FJ750950. The longth of complete sequence is 5003bp,contain an open reading frame which codes a polypeptide of 362 amino acids.The predicted nucleotide sequence similarity with human,mouse,cattle,horse,dog and macaca mulatta FP gene were respectively 77%,81%,81%,80%,89%,74%and the amino acid sequence were respectively 66.3%, 82.8%,87.6%,86.4%,86.6%,85.9%.There were a G-protein linked receptor family at 43-304 position(Prosite:PS50262,Score20.693).IMpRH was used for determining the chromosomal regional location of the pig FP gene,The result of the data analysis indicated that FP gene was mapped to the region of SSC6q31-q32,with close linkage to SW16472(LOD=12.61)and S0299(LOD=10.56),and normal linkage to SWR726 (LOD=6.79).Result of RT-PCT revealed that FP gene was highly expressed in ovary and endometrium,followed by spleen,heart,kidney,stomach,and no expressed in liver.(3) The full-length cDNA sequence from the porcine CO986776 was cloned.The sequences have been deposited in GenBank database,the accession number is FJ603023. The longth of complete sequence is 2585bp,contain an open reading frame which is codes a polypeptide of 176 amino acids.The predicted nucleotide sequence similarity with human,mouse,cattle and macaca mulatta MAL2 gene were respectively75%, 70%,81%,74%and the amino acid sequence were respectively90%,90%,93%,84%. Constructive domain analysis by ExPASy ScanProsite indicate that there is a MARVEL domain(Prosite:PS51225,Score 15.143) at 31-175 position.The result of the data analysis indicated that MAL2 gene was mapped to the region of SSC4p12-p¹³, with close linkage to SWR73(LOD=6.38) and normal linkage to S0001(LOD=6.35) and SWR2179(LOD=5.07).Result of RT-PCT revealed that MAL2 gene was highly expressed in stomach,followed by liver,kidney,endometrium,ovary,and no expressed in heart and spleen.