Grant D F, Spearow J L, Storms D H, Edelhoff S, Adler D A, Disteche C M, Taylor B A, Hammock B D
Department of Entomology and Environmental Toxicology, University of California, Davis, CA 95616.
Pharmacogenetics. 1994 Apr;4(2):64-72. doi: 10.1097/00008571-199404000-00003.
The chromosomal location of a murine soluble epoxide hydrolase gene was determined using in situ mapping, restriction fragment length polymorphism (RFLP) and simple sequence length polymorphism (SSLP) analysis. In situ hybridization to mouse metaphase chromosomes using a soluble epoxide hydrolase cDNA probe showed that soluble epoxide hydrolase maps at band D of chromosome 14. An RFLP found between Mus castaneus (CAST) and Mus musculus (MEV) was used to map the soluble epoxide hydrolase gene in CAST x MEV intersubspecific testcross progeny to 14 cM from the Np-1 locus on mouse chromosome 14. SSLP markers were then used to confirm the location of soluble epoxide hydrolase at 14.0 +/- 3.7 cM distal to Np-1 and 19.2 +/- 4.3 cM proximal to D14Mit7. This region of mouse chromosome 14 is homologous with human chromosomes 8, 13 and 14. Enzyme assays and immunoblotting results suggest significant quantitative differences in expression of soluble epoxide hydrolase among three mouse strains. Northern blotting analysis showed that soluble epoxide hydrolase mRNA levels were correlated with the relative level of soluble epoxide hydrolase enzyme activity and soluble epoxide hydrolase protein in all three mouse strains.
利用原位定位、限制性片段长度多态性(RFLP)和简单序列长度多态性(SSLP)分析确定了小鼠可溶性环氧化物水解酶基因的染色体定位。使用可溶性环氧化物水解酶cDNA探针与小鼠中期染色体进行原位杂交,结果显示可溶性环氧化物水解酶定位于14号染色体的D带。利用在栗色小鼠(CAST)和小家鼠(MEV)之间发现的一种RFLP,将CAST×MEV种间回交后代中的可溶性环氧化物水解酶基因定位到小鼠14号染色体上距Np-1基因座14 cM处。随后使用SSLP标记确认可溶性环氧化物水解酶位于距Np-1远端14.0±3.7 cM以及距D14Mit7近端19.2±4.3 cM处。小鼠14号染色体的这一区域与人类8号、13号和14号染色体同源。酶活性测定和免疫印迹结果表明,三种小鼠品系中可溶性环氧化物水解酶的表达存在显著的数量差异。Northern印迹分析表明,在所有三种小鼠品系中,可溶性环氧化物水解酶mRNA水平与可溶性环氧化物水解酶活性和可溶性环氧化物水解酶蛋白的相对水平相关。