Expression Of ADSL Gene In Xupu Geese And Landes Geese And The Association Reaseach Between SNPs And Inosine Monophosphate Content

The c DNA rapid amplification(RACE)technique was used in this study to clone the c DNA full length of adenosine succinic acid lyase(ADSL)gene of Xupu geese,and made bioinformatics analysis of its nucleic acid and proteins sequences.The distribution rules of inosine monophosphate(IMP)in different tissues of Xupu geese and Landes geese was compared by high performance liquid chromatography(HPLC)technique,and the expression level of ADSL gene in different breeds and tissues in Xupu geese and Landes geese was detected.The expression difference of ADSL gene in different breeds of geese tissues was detected by real-time fluorescent quantitative PCR.The polymorphisms of ADSL gene between Xupu geese and Landes geese was compared by direct sequencing to provide reference for the development and breeding of high quality meat geese.The main results were as follows:1.Cloning and bioinformatics analysis of ADSL gene.The full length c DNA sequence of ADSL gene in Xupu geese was 1496 bp,containing a202 bp 5’UTR,a 31 bp 3’ UTR and a 1263 bp open reading frame(ORF),and encoding a protein of 420 amino acide,no signal peptide sites,non-secreted protein(Gen Bank accession number: MK161097).It was predicted by subcellular localization that the gene was mainly present in the cytoplasm and was an unstable hydrophilic protein.The ADSL gene sequences of 11 species were downloaded from Genebank database for homology analysis,the Xupu geese ADSL showed 99.68%,88.35%,85.64%,82.24% amino acid sequence identities with previously described homologues from Zhedong White geese(Anser cygonodes domesticus),duck(Anas platyrhynchos),chicken(Gallus gallus),pigceon(Columba livia),respectively.Based on the evolutionary relationship of the gene,the Xupu Geese was the most related to the Zhedong White geese,followed by ducks and chickens,which was far related to mammals.2.The differences of IMP in different and breeds and tissues.The average IMP contents of breast and leg muscles in Xupu geese respectively were2.61±0.46 mg/g and 2.15±0.61 mg/g.The average IMP contents of breast and leg muscles in Landes geese were respectively 2.34±0.56 mg/kg and 1.42±0.61 mg/g,respectively.According to IMP content analysis showed that IMP content in breast muscle was extreme significantly higher than that in leg muscle in different varieties(P<0.01).In addition,the IMP content of breast muscle in Xupu geese was extreme significantly higher than that of the Landes geese breast and leg muscles(P<0.01).IMP content in leg muscle of Xupu geese was extreme significantly higher than that of Landes geese(P<0.01),the breast muscle of Landes geese was highter than the leg muscle of Xupu geese,but there was not significant difference(P>0.05).3.The expression level of ADSL gene in different breeds and tissues of goose.ADSL gene was expressed in five tissues of Xupu geese.The expression levels of ADSL gene in Xupu geese were successively from high to low: breast muscle,leg muscle,liver,sebum and abdominal fat.The expression level of ADSL gene in breast muscle was significantly higher than that in sebum and abdominal fat(P< 0.05),but there was no significant difference in other tissues(P>0.05).The expression of ADSL gene in Landes geese from high to low was: breast muscle,leg muscle,abdominal fat,sebum,and liver.There was no significant difference in gene expression between breast muscle and leg muscle(P>0.05).The expression of ADSL gene in muscle breast and leg muscle were extreme significantly higher than abdominal fat,sebum,and liver(P<0.01),and there was no significant difference among abdominal fat and sebum and liver(P>0.05).4.SNPs screening of ADSL geneThe single nucleotide polymorphisms(SNPs)of ADSL gene in Xupu geese and Landes geese were screened by direct sequencing.And 7 and 13 mutation sites were found in Xupu geese and Landes geese,respectively.Among them,there was 5 mutation sites in Xupu geese that meet the Hardy-Weinberg equilibrium,and 13 mutation sites in Landes geese was consistent with Hardy-Weinberg equilibrium.In addition,the correlation analysis between ADSL gene mutation sites and IMP content showed that C-10793 G,A-10927 G and T-11033 G were significantly associated with IMP content in the breast muscle(P<0.05).While no mutation site associated with IMP content is found in Landes geese.In conclusion,IMP content was significantly different in different breeds and tissues of geese.And as the main candidate gene affecting IMP deposition,ADSL gene affects the synthesis of IMP in the vivo.In this study,the SNPs sites were found to be significantly correlated with IMP content,which provides a theoretical basis for the development of geese breed resources.