苹果酸酶探针可检测到与编码其他代谢酶的基因座紧密连锁的交叉杂交序列。

A malic enzyme probe detects cross-hybridizing sequences closely linked to loci encoding other metabolic enzymes.

作者信息

Cobb R R, Rinchik E, Barnett L B, Burkhart J G, Lewis S E

机构信息

Chemistry and Life Sciences Group, Research Triangle Institute, Research Triangle Park, North Carolina 27709.

出版信息

Genomics. 1992 Feb;12(2):405-8. doi: 10.1016/0888-7543(92)90393-7.

DOI:10.1016/0888-7543(92)90393-7

PMID:1346782

Abstract

The cytoplasmic malic enzyme (Mod-1) catalyzes the oxidative decarboxylation of malate: malate + NADP+----pyruvate + CO2 + NADPH + H+. Using a cDNA clone of Mod-1 as a probe, two new DNA markers not at the Mod-1 locus (restriction fragment length polymorphisms, RFLP) were detected by Southern blot analysis that showed extensive homology to Mod-1 sequences. Linkage of each restriction fragment length polymorphism to loci other than Mod-1 was assessed using the BXD (C57BL/6J x DBA/2J) recombinant inbred strains and confirmed by backcrosses. One polymorphic site, designated D9Rti1, was found to be closely linked to the phosphoglucomutase (Pgm-3) locus on Chromosome 9. The other hybridization site, designated D1Rti2, was closely linked to the isocitrate dehydrogenase (Idh-1) locus on Chromosome 1. The data presented imply that Mod-1 homologous sequences are tightly linked to three different metabolic enzymes.

摘要

细胞质苹果酸酶（Mod-1）催化苹果酸的氧化脱羧反应：苹果酸 + NADP⁺ → 丙酮酸 + CO₂ + NADPH + H⁺。以Mod-1的cDNA克隆作为探针，通过Southern印迹分析检测到两个不在Mod-1基因座的新DNA标记（限制性片段长度多态性，RFLP），它们与Mod-1序列具有广泛的同源性。使用BXD（C57BL/6J×DBA/2J）重组近交系评估每个限制性片段长度多态性与Mod-1以外基因座的连锁关系，并通过回交进行确认。发现一个多态性位点，命名为D9Rti1，与9号染色体上的磷酸葡萄糖变位酶（Pgm-3）基因座紧密连锁。另一个杂交位点，命名为D1Rti2，与1号染色体上的异柠檬酸脱氢酶（Idh-1）基因座紧密连锁。所呈现的数据表明，Mod-1同源序列与三种不同的代谢酶紧密连锁。