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一种用于检测和测量不同物种棕色脂肪组织中解偶联蛋白mRNA的27聚体寡核苷酸探针。

A 27-mer oligonucleotide probe for the detection and measurement of the mRNA for uncoupling protein in brown adipose tissue of different species.

作者信息

Brander F, Keith J S, Trayhurn P

机构信息

Division of Biochemical Sciences, Rowett Research Institute, Bucksburn, Aberdeen, Scotland, U.K.

出版信息

Comp Biochem Physiol B. 1993 Jan;104(1):125-31. doi: 10.1016/0305-0491(93)90348-9.

DOI:10.1016/0305-0491(93)90348-9
PMID:8448986
Abstract
  1. Data from rats, cattle, mice, rabbits and humans indicate considerable species heterogeneity in the sequence of the gene coding for the mitochondrial uncoupling protein (UCP) in brown adipose tissue. A 27-base sequence of an exon region of the gene is, however, identical in rats and cattle; in mice, rabbits, and humans this same region shows only a single base difference from the sequence in rats and cattle. 2. A 27-mer oligonucleotide (3'-TGGAAGGGCGACCTGTGGCGGTTTCAG-5') complementary to the conserved region of the rat and cattle UCP genes has been synthesized as a potential probe for UCP mRNA in widely differing species. 3. Northern blots of RNA from rat brown fat showed that the oligonucleotide hybridized with a 1.5 kbase mRNA, indicative of UCP mRNA. No hybridization was observed with RNA from white fat (subcutaneous, internal), liver, kidney, skeletal muscle, heart and brain. Acute cold-exposure of rats and mice led to an increase in UCP mRNA level, while streptozotocin-induced diabetes resulted in a decrease. 4. The oligonucleotide hybridized with a 1.5 (or 1.9) kbase mRNA from brown fat of rats, mice, golden hamsters, Djungarian hamsters, and newborn rabbits, pipistrelle bats, lambs, goats and red deer. 5. The 27-mer oligonucleotide provides a simple probe for UCP mRNA across a wide range of mammals, obviating any need to obtain species-specific cDNAs.
摘要
  1. 来自大鼠、牛、小鼠、兔子和人类的数据表明,棕色脂肪组织中线粒体解偶联蛋白(UCP)编码基因的序列存在相当大的物种异质性。然而,该基因一个外显子区域的27个碱基序列在大鼠和牛中是相同的;在小鼠、兔子和人类中,这个相同区域与大鼠和牛的序列仅存在一个碱基差异。2. 已合成了一种与大鼠和牛UCP基因保守区域互补的27聚体寡核苷酸(3'-TGGAAGGGCGACCTGTGGCGGTTTCAG-5'),作为一种潜在的探针,用于检测广泛不同物种中的UCP mRNA。3. 大鼠棕色脂肪RNA的Northern印迹显示,该寡核苷酸与一个1.5千碱基的mRNA杂交,表明是UCP mRNA。未观察到与白色脂肪(皮下、内脏)、肝脏、肾脏、骨骼肌、心脏和大脑的RNA发生杂交。对大鼠和小鼠进行急性冷暴露会导致UCP mRNA水平升高,而链脲佐菌素诱导的糖尿病则会使其降低。4. 该寡核苷酸与大鼠、小鼠、金黄仓鼠、黑线仓鼠、新生兔子、伏翼蝙蝠、羔羊、山羊和马鹿棕色脂肪中的1.5(或1.9)千碱基mRNA杂交。5. 这种27聚体寡核苷酸为广泛的哺乳动物中的UCP mRNA提供了一种简单的探针,无需获取物种特异性的cDNA。

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A 27-mer oligonucleotide probe for the detection and measurement of the mRNA for uncoupling protein in brown adipose tissue of different species.一种用于检测和测量不同物种棕色脂肪组织中解偶联蛋白mRNA的27聚体寡核苷酸探针。
Comp Biochem Physiol B. 1993 Jan;104(1):125-31. doi: 10.1016/0305-0491(93)90348-9.
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Rapid chemiluminescent detection of the mRNA for uncoupling protein in brown adipose tissue by northern hybridization with a 32-mer oligonucleotide end-labelled with digoxigenin.通过与用地高辛配基末端标记的32聚体寡核苷酸进行Northern杂交,对棕色脂肪组织中解偶联蛋白的mRNA进行快速化学发光检测。
Int J Obes Relat Metab Disord. 1994 Jun;18(6):449-52.
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Postnatal development of uncoupling protein, uncoupling protein mRNA, and GLUT4 in adipose tissues of goats.山羊脂肪组织中解偶联蛋白、解偶联蛋白mRNA及葡萄糖转运蛋白4的出生后发育
Am J Physiol. 1993 Sep;265(3 Pt 2):R676-82. doi: 10.1152/ajpregu.1993.265.3.R676.
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Oligonucleotide probe for the cross-species measurement of the mRNA for uncoupling protein in brown adipose tissue.用于跨物种测量棕色脂肪组织中解偶联蛋白mRNA的寡核苷酸探针。
Biochem Soc Trans. 1990 Dec;18(6):1261. doi: 10.1042/bst0181261.
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Sequential changes in the expression of mitochondrial protein mRNA during the development of brown adipose tissue in bovine and ovine species. Sudden occurrence of uncoupling protein mRNA during embryogenesis and its disappearance after birth.牛和羊物种棕色脂肪组织发育过程中线粒体蛋白mRNA表达的顺序变化。胚胎发生过程中解偶联蛋白mRNA的突然出现及其出生后的消失。
Biochem J. 1989 Feb 1;257(3):665-71. doi: 10.1042/bj2570665.
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Molecular approach to thermogenesis in brown adipose tissue: cDNA cloning of the mitochondrial uncoupling protein.棕色脂肪组织产热的分子机制:线粒体解偶联蛋白的cDNA克隆
Proc Natl Acad Sci U S A. 1985 Jan;82(2):445-8. doi: 10.1073/pnas.82.2.445.
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Detection of brown adipose tissue uncoupling protein mRNA in adult patients by a human genomic probe.利用人类基因组探针检测成年患者棕色脂肪组织解偶联蛋白mRNA
Clin Sci (Lond). 1988 Jul;75(1):21-7. doi: 10.1042/cs0750021.
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Chemiluminescent detection of uncoupling protein mRNA on northern blots with a digoxigenin (DIG) end-labelled oligonucleotide.
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The beta 3-adrenoceptor agonist ICI-D7114 is not as efficient on reinduction of uncoupling protein mRNA in sheep as it is in dogs and smaller species.β3-肾上腺素能受体激动剂ICI-D7114在诱导绵羊解偶联蛋白信使核糖核酸重新表达方面的效果不如在犬类和较小物种中那样显著。
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Rabbit brown adipose tissue uncoupling protein MRNA: use of only one of two polyadenylation signals in its processing.兔棕色脂肪组织解偶联蛋白信使核糖核酸:在其加工过程中仅使用两个聚腺苷酸化信号之一。
Biochem Biophys Res Commun. 1989 May 30;161(1):156-61. doi: 10.1016/0006-291x(89)91574-x.

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