cDNA Cloning and Characterization of Adipose Triglyceride Lipase Gene in Zebra Finch ( Taeniopygia guttata ) and Java Sparrow ( Padda oryzivora )
Abstract
Adipose triglyceride lipase (ATGL) is a crucial lipase that catalyzes triglyceride hydrolysis. In this study, reverse transcription (RT)-polymerase chain reaction, rapid amplification of cDNA ends, and mRNA real-time analysis were performed to characterize the ATGL gene in zebra finch and Java sparrow. Two cDNA of the zebra finch ATGL ( zfATGL ) gene were 1877 and 1626 bp were obtained; both encoded the same 481 amino acid protein. A 251 bp indel in the 3′ UTR led to two variant transcripts. The zfATGL gene at chromosome 5 spanned over 28 Kb in length and comprised 9 exons. As in chicken, both the zfATGL gene and its intron 1 were much longer than mammals, which seemed to be one characteristic of ATGL gene in birds. The zfATGL gene was predominantly expressed in abdominal fat, subcutaneous fat, breast muscle, and leg muscle in both male and female. Two cDNA (1938 and 1885 bp) of the Java sparrow ATGL gene were also obtained, both of which encoded the same 481 amino acid protein. A 53 bp indel in 3′ UTR led to two variant transcripts. The zfATGL gene was predominantly expressed in breast muscle, subcutaneous fat, abdominal fat, heart, and leg muscle. ATGL homology analysis showed that GASAG and GCGFLG motifs in the patatin region were conserved among 16 species. It was further indicated that the 251- and 53 bp indels at the 3′ UTR of ATGL were potential binding sites for miRNA tgu-miR-2987 . In conclusion, our study identified the full length cDNA and their variant transcripts of zfATGL and Java sparrow ATGL , and they were predominantly expressed in muscle, fat, and heart tissues.