Cloning and expression of a chicken alpha-amylase gene.
We have isolated and sequenced a genomic clone for a pancreatic alpha-amylase gene (amy) of the chicken (Gallus gallus). The gene is interrupted by nine introns, spans over 4 kb, and encodes a protein (AMY) of 512 aa that is 83% identical to the human pancreatic alpha-amylase enzyme. Southern blot analysis of chicken DNA revealed two distinct pancreatic amy loci. In addition, we have generated a cDNA from chicken pancreatic RNA corresponding to the coding sequence of the genomic clone. The cDNA was inserted into a yeast expression vector, and the resulting construct used to transform Saccharomyces cerevisiae cells. Transformed yeast cells synthesized and secreted active AMY enzyme, and the gel migration pattern of the alpha-amylase produced by the yeast cells was identical to that of the native chicken enzyme.
Benkel
B F
BF
Agriculture and Agri-Food Canada, Centre for Food and Animal Research, Ottawa, Ont.
Nguyen
T
T
Ahluwalia
N
N
Benkel
K I
KI
Hickey
D A
DA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002645:Chickens; D003001:Cloning, Molecular; D015971:Gene Expression Regulation, Enzymologic; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D000516:alpha-Amylases / Q000235:genetics*
D012333:RNA, Messenger; D000516:alpha-Amylases