Cloning, Tissue Expression And Genetic Variation Of Bovine ATGL Gene

In this study, reverse transcriptase polymerase chain reaction (RT-PCR), semi-quantitative RT-PCR, pyrosequencing methods were employed to obtain the complete CDSsequence of ATGL,the tissue expression of ATGL and screen for the genetic variation in ATGL gene among 702 individuals of five breeds. Association analysis between ATGL gene polymorphisms and some economic traits in Qinchuan populations was studyed.The main results were as follows:1. The CDS of ATGL gene was gained (FJ798978). The full length of the CDS was 1461 bp, which encoded a polypeptide of 486 amino acids. The ATGL protein contains one Patatin domains and was comprised of fiveαhelix and sixβ-sheet. The homologies of cDNA nucleotide sequence of ATGL gene with the counterparts of homo, mouse, rat, dog and swine were 87.3%, 84.7% , 82.4%, 86.8%, 87.5%. Phylogenetic tree analysis showed that the cattle and pigs had the closest relationship.2. Semi-quantitative RT-PCR was used to detect the mRNA expression of bovine ATGL gene. The results showed that ATGL gene was highly expressed in fat and rumenbut was not detected in testicle, indicating that the ATGL gene in various tissues of cattle plays a wide range of biological effects.3. Based on the pyrosequencing technology, four novel missense mutations were indentified, which were 3166T>C, 3289G>C, 3514A>T and 3947G>T, respectively. The 3166T>C locus was significantly associated with backfat thickness (BFT) and marbling score (MS). Cattle with genotype CC have higher BFT and MS than those with genotype TT. The 3289G>C mutation had significant effect on backfat thickness (BFT), cattle with genotype CC had higher BFT than those with genotype GG and GC. The 3514A>T locus was significantly associated with dressing percentage (DP) and MS. Cattle with genotype TT had higher DP and MS than those with genotype AA and AT. The 3947G>T mutation was significantly associated with MS and Meat tenderness (MT). Individuals with genotype TT and TG had higher MS than those with genotype GG. The meat of individuals with genotype GG and GT was more tender than those with genotype TT. The results suggested that the four SNPs of the ATGL gene may be useful as genetic markers for carcass and meat quality traits in cattle.