Pemble S E, Taylor J B, Craig R K, Ketterer B
Biochem J. 1986 Sep 1;238(2):373-8. doi: 10.1042/bj2380373.
The content of GSH transferase mRNAs in poly(A)-containing RNA isolated from eight rat tissues was examined by immunoprecipitation of cell-free translation products and by Northern blotting. Considerable tissue-specific distribution and heterogeneity of immunoprecipitable GSH transferase subunits 1 and 2 synthesized in vitro was observed. These results were confirmed by Northern blotting using a 32P-labelled subunit 1 cDNA probe. The same probe, used in a Southern blot analysis of genomic DNA, provided confirmation that GSH transferase subunits 1 and 2 comprise a multigenic family in the rat. The results show that the selection of cDNA clones coding for chosen subunits can be made easier by making use of qualitative and quantitative tissue differences in GSH transferase mRNAs.
通过对无细胞翻译产物进行免疫沉淀以及Northern印迹法,检测了从8种大鼠组织中分离出的含多聚腺苷酸(poly(A))RNA中谷胱甘肽S-转移酶(GSH transferase)mRNA的含量。观察到体外合成的可免疫沉淀的GSH转移酶亚基1和2具有明显的组织特异性分布和异质性。使用32P标记的亚基1 cDNA探针进行Northern印迹法证实了这些结果。在对基因组DNA进行Southern印迹分析时使用相同的探针,进一步证实了GSH转移酶亚基1和2在大鼠中构成一个多基因家族。结果表明,利用GSH转移酶mRNA在质量和数量上的组织差异,能够更轻松地选择编码特定亚基的cDNA克隆。