Comparison of amylase mRNAs from rat parotid gland, pancreas and liver using reverse transcriptase-polymerase chain reaction.
The expression of mRNA for amylase was examined using the reverse transcriptase-polymerase chain reaction (RT-PCR). An amylase product was strongly detected in parotid and pancreas, but less strongly in liver. The degree of identity between the PCR products was assessed by restriction-enzyme mapping using two restriction enzymes, EcoRI and ScaI, and DNA sequencing. The PCR product from pancreas was cut by both EcoRI and ScaI, while the products from parotid and liver were cut by EcoRI but not by ScaI. The sequence of the parotid product was 90.4% homologous to that of the pancreas, and 100% homologous to that of the liver. These results indicate that the same amylase mRNA may be expressed in parotid and liver. In addition, the expression of amylase mRNAs in other rat tissues was investigated using RT-PCR, and the sensitivity of each PCR product to ScaI was tested. A weak single band was detected in submandibular gland, sublingual gland and stomach. ScaI digestion cut the stomach product into two fragments, but had no effect on the submandibular and sublingual products. Thus, it may be possible to classify amylase isoenzymes into pancreatic and parotid types based on the sensitivity of their PCR products to ScaI.
Nezu
Akihiro
A
Department of Dental Pharmacology, School of Dentistry, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido 061-0293, Japan. a-nezu@hoku-iryo-u-ac.jp
Morita
Takao
T
Tanimura
Akihiko
A
Tojyo
Yosuke
Y
D000681:Amylases / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D015246:Deoxyribonuclease EcoRI; D015252:Deoxyribonucleases, Type II Site-Specific; D007527:Isoenzymes; D008099:Liver / Q000201:enzymology*; D008297:Male; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology*; D010306:Parotid Gland / Q000201:enzymology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D015183:Restriction Mapping; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid
D007527:Isoenzymes; D012333:RNA, Messenger; D015246:Deoxyribonuclease EcoRI; C047020:endodeoxyribonuclease ScaI; D015252:Deoxyribonucleases, Type II Site-Specific; D000681:Amylases