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小鼠sn-甘油-3-磷酸脱氢酶:cDNA序列的分子克隆与基因定位

Mouse sn-glycerol-3-phosphate dehydrogenase: molecular cloning and genetic mapping of a cDNA sequence.

作者信息

Kozak L P, Birkenmeier E H

出版信息

Proc Natl Acad Sci U S A. 1983 May;80(10):3020-4. doi: 10.1073/pnas.80.10.3020.

Abstract

The isozymes of glycerol-3-phosphate dehydrogenase (GPDH; sn-glycerol-3-phosphate:NAD+2-oxidoreductase, EC 1.1.1.8) in tissues of the mouse are coded for by two structural genes, Gdc-1 and Gdc-2, located on chromosomes 15 and 9, respectively. In order to investigate the expression of these genes, we isolated a GPDH cDNA clone from a mRNA preparation isolated from brown adipose tissue. The GPDH cDNA clone was identified by colony hybridization and hybrid selection of a mRNA that was translated in vitro to produce immunoprecipitable GPDH protein. In blot analysis, the GPDH cDNA hybridized to a single mRNA species that migrated at the position of 23S ribosomal RNA. This GPDH cDNA clone was mapped to the Gdc-1 locus by identification of a restriction enzyme polymorphism present in genomic DNA isolated from Gdc-1 congeneic lines of mice.

摘要

小鼠组织中甘油-3-磷酸脱氢酶(GPDH;sn-甘油-3-磷酸:NAD+ 2-氧化还原酶,EC 1.1.1.8)的同工酶由分别位于15号和9号染色体上的两个结构基因Gdc-1和Gdc-2编码。为了研究这些基因的表达,我们从棕色脂肪组织分离的mRNA制剂中分离出一个GPDH cDNA克隆。通过菌落杂交和对体外翻译产生可免疫沉淀的GPDH蛋白的mRNA进行杂交筛选来鉴定GPDH cDNA克隆。在印迹分析中,GPDH cDNA与迁移到23S核糖体RNA位置的单一mRNA物种杂交。通过鉴定从小鼠Gdc-1同源系分离的基因组DNA中存在的限制性酶多态性,将该GPDH cDNA克隆定位到Gdc-1基因座。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f5b/393965/dccbfc9a30b1/pnas00636-0220-a.jpg

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