Abstract
Portions of at least nine distinct rat amylase genes or pseudogenes have been isolated. Cloned rat genomic DNA fragments containing complete or major portions of seven of these have been examined by heteroduplex analysis and fall within two separate groups based on their degree of homology. Four gene sequences comprising one of these groups are closely related to pancreatic amylase mRNA. The other group shows significant nonhomology to both pancreatic and parotid amylase cDNAs and may represent an additional gene type(s). All of the cloned amylase gene sequences are found in rat genomic DNA. Additional amylase sequences which have not yet been cloned are also detected. Comparison of DNA from individual Sprague-Dawley rats by Southern blotting techniques indicates allelic variation at multiple amylase loci.
Highlights
RESULTSTions of amylase gene sequences and extend in either the 5' or 3' direction. A more detailed restriction endonuclease analysis
Screening of a Rat Genomic DNA-A Charon 4A DNA Library for DNA fragments containing complete or major portionAs mylase Gene Sequences-A Sprague-Dawley rat genomic DNAA of seven of these have been examined by heteroduplex Charon 4A library [9] was screened for amylase gene sequences as analysis and fall within two separate groups based on described by Benton and Davis [10]
Cloned genomic DNA fragments were digested with foundinratgenomicDNA.Additionalamylase se- restriction endonucleases, subjected to electrophoresis on 1%agarose quences which have not yet been cloned are de- gels, andtransferred to nitrocellulose filters
Summary
Tions of amylase gene sequences and extend in either the 5' or 3' direction. A more detailed restriction endonuclease analysis. Cloned Amylase Gene Sequences inR a t Genomic DNAFig. 2 shows a Southern blot of EcoRI-digested liver DNA from 10 individual Sprague-Dawleyrats hybridized with either the 5' (Fig. 2 A ) or 3' (Fig. 2B) cDNA probe. All of the EcoRI fragments from the cloned amylase genes (Fig. 1) are observed except the 0.2-kb fragment from Amy 5 We believe this results froma low level hybridization. In South- Fig. 2 and TableI reveal EcoRI fragmentosf molecular ern blots of cloned Amy 5 DNA the0.2-kb fragment is detected length 15kb, 4.4 kb, and 4.2 kb (hybridizing with the5' probe) but exhibitsa low intensity hybridization band relative to the and 15kb, 6.0 kb, 2.6 kb, 1.25 kb, and0.4 kb
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