Jacobsson A, Stadler U, Glotzer M A, Kozak L P
J Biol Chem. 1985 Dec 25;260(30):16250-4.
We have identified cDNAs clones for several cold-inducible mRNAs from the brown adipose tissue of mice. pCIN-1, a plasmid with a 900-base pair insert, encoded the mitochondrial uncoupling protein (UCP) as determined by the ability of the cDNA insert to select, by hybridization, an mRNA that could be translated into a 32,000-Da protein immunoprecipitable with anti-UCP antibodies. Nine tissues were analyzed; however, UCP cDNA hybridized to an mRNA species of 1.6 and 2.0 kilobase pairs only in brown adipose tissue. A maximum induction of 10-fold occurred within 6 h of exposure to cold (5 degrees C). A BamHI restriction fragment polymorphism detected by Southern blot analysis of genomic DNA in recombinant inbred mouse strains allowed us to map the UCP gene to Chromosome 8. The analysis of the UCP gene expression in diabetic (db) and obese (ob) mice maintained at 27 degrees C for 3 days followed by cold exposure for 4 h at 5 degrees C indicated that UCP mRNA levels in mutant mice were unaffected at 27 degrees C and only slightly reduced at 5 degrees C. Accordingly, the inability of diabetic and obese mice to thermoregulate is not associated with a lack of UCP mRNA induction.
我们已经从小鼠棕色脂肪组织中鉴定出了几种冷诱导mRNA的cDNA克隆。pCIN-1是一种带有900个碱基对插入片段的质粒，通过cDNA插入片段能够通过杂交选择出一种mRNA，该mRNA可被翻译成一种能与抗UCP抗体发生免疫沉淀反应的32,000道尔顿蛋白质，从而确定其编码线粒体解偶联蛋白（UCP）。我们分析了9种组织；然而，UCP cDNA仅在棕色脂肪组织中与1.6和2.0千碱基对的mRNA种类杂交。在暴露于寒冷（5摄氏度）6小时内，诱导倍数最高可达10倍。通过对重组近交系小鼠品系的基因组DNA进行Southern印迹分析检测到的BamHI限制性片段多态性，使我们能够将UCP基因定位到8号染色体上。对在27摄氏度饲养3天，随后在5摄氏度冷暴露4小时的糖尿病（db）和肥胖（ob）小鼠的UCP基因表达分析表明，突变小鼠中的UCP mRNA水平在27摄氏度时未受影响，在5摄氏度时仅略有降低。因此，糖尿病和肥胖小鼠无法进行体温调节与缺乏UCP mRNA诱导无关。
