Mocharla H, Mocharla R, Hodes M E
Department of Medical Genetics, School of Medicine, Indiana University, Indianapolis 46202-5251.
Nucleic Acids Res. 1990 Feb 25;18(4):1031-6. doi: 10.1093/nar/18.4.1031.
We studied the distribution of alpha-amylase mRNA in normal dog tissues by northern blotting (NB) and reverse transcription-polymerase chain reaction (RT-PCR) with human pancreatic (AMY2) and salivary (AMY1) alpha-amylase cDNA-specific primers. Analysis of poly(A+) RNA from various normal tissues by NB indicated the presence of detectable levels of alpha-amylase mRNA transcripts only in pancreas. Dot-blot analysis of DNA amplified with primers common to both (human) isoamylase mRNAs showed presence of alpha-amylase gene transcripts not only in pancreas but also in liver, small intestine, large intestine and fallopian tube. Traces of amylase gene transcripts were also observed in ovary, uterus and lung. Interestingly, amylase transcripts were not detectable in the parotid gland by NB or RT-PCR. We have also localized alpha-amylase mRNA transcripts to dog pancreas by in situ transcription and in situ hybridization. Our results suggest that there is high degree of homology between the alpha-amylase mRNA sequences in dog and human at least in the exon 3-4 regions of the human gene.
我们通过Northern印迹法(NB)以及使用人胰腺(AMY2)和唾液(AMY1)α-淀粉酶cDNA特异性引物的逆转录聚合酶链反应(RT-PCR),研究了α-淀粉酶mRNA在正常犬组织中的分布。通过NB对来自各种正常组织的聚腺苷酸(A+)RNA进行分析,结果表明仅在胰腺中存在可检测水平的α-淀粉酶mRNA转录本。用两种(人)同工淀粉酶mRNA共有的引物扩增DNA的点印迹分析显示,α-淀粉酶基因转录本不仅存在于胰腺中,还存在于肝脏、小肠、大肠和输卵管中。在卵巢、子宫和肺中也观察到痕量的淀粉酶基因转录本。有趣的是,通过NB或RT-PCR在腮腺中均未检测到淀粉酶转录本。我们还通过原位转录和原位杂交将α-淀粉酶mRNA转录本定位到犬胰腺。我们的结果表明,犬和人α-淀粉酶mRNA序列之间至少在人基因的外显子3-4区域具有高度同源性。