Shen P, Howlett G J
Russell Grimwade School of Biochemistry, University of Melbourne, Parkville, Victoria, Australia.
Arch Biochem Biophys. 1992 Sep;297(2):345-53. doi: 10.1016/0003-9861(92)90683-n.
Restriction fragments isolated from a 17-kb rat genomic DNA clone containing the gene for apolipoprotein (apo) E were radiolabeled and used to screen a rat liver cDNA library. A cDNA clone hybridizing to a 6-kb genomic DNA fragment was isolated and the nucleotide sequence of the cDNA insert determined. The sequence was homologous to the sequence for human apo C-I and was used to derive the corresponding amino acid sequence. Unlike human apo C-I, mature rat apo C-I contains histidine, lacks valine, and has alanine at the C terminus and aspartate as the N terminus. Screening the rat liver cDNA library with a radiolabeled 1.9-kb restriction fragment from the genomic DNA clone containing the rat apo E gene identified another cDNA clone (ECL cDNA). Nucleotide sequencing yielded a derived 75-amino-acid sequence for the ECL protein with a hydrophobicity profile similar to that of rat apo C-I. Northern analysis demonstrated a 0.50-kb band for ECL mRNA. The tissue-specific expression of the gene is similar to that of rat apo C-I. This study indicates that the rat apo C-I and ECL genes are closely linked, about 4.5 and 12 kb downstream of the apo E gene, respectively.
从一个包含载脂蛋白(apo)E基因的17kb大鼠基因组DNA克隆中分离出的限制性片段经放射性标记后,用于筛选大鼠肝脏cDNA文库。分离出一个与6kb基因组DNA片段杂交的cDNA克隆,并测定了该cDNA插入片段的核苷酸序列。该序列与人apo C-I的序列同源,并用于推导相应的氨基酸序列。与人类apo C-I不同,成熟的大鼠apo C-I含有组氨酸,缺乏缬氨酸,C末端为丙氨酸,N末端为天冬氨酸。用来自包含大鼠apo E基因的基因组DNA克隆的放射性标记的1.9kb限制性片段筛选大鼠肝脏cDNA文库,鉴定出另一个cDNA克隆(ECL cDNA)。核苷酸测序得出了ECL蛋白的75个氨基酸的推导序列,其疏水性图谱与大鼠apo C-I相似。Northern分析显示ECL mRNA有一条0.50kb的条带。该基因的组织特异性表达与大鼠apo C-I相似。这项研究表明,大鼠apo C-I和ECL基因紧密相连,分别位于apo E基因下游约4.5kb和12kb处。