编码人酸性鞘磷脂酶的cDNA克隆的分离:发生交替处理的转录本。

文章的细节

引用

Quintern LE, Schuchman EH, Levran O, Suchi M, Ferlinz K, Reinke H, Sandhoff K, Desnick RJ

编码人酸性鞘磷脂酶的cDNA克隆的分离:发生交替处理的转录本。

EMBO J. 1989 9月8日(9):2469-73。

PubMed ID
2555181 (PubMed视图
摘要

从人尿液中纯化了酸性鞘磷脂酶(sphingomyelin phosphodiesterase, EC 3.1.4.12),并对12个胰蛋白酶肽进行了微测序(128个残基)。根据密码子冗余度最小的区域,合成了4种寡核苷酸混合物,其中寡核苷酸混合物1 (20mer;256 mix)用于从人成纤维细胞cDNA文库中筛选3 X 10(6)个独立的重组子。推测阳性克隆92个,用寡核苷酸混合物2-4进行Southern杂交分析。这些研究揭示了两组克隆;第一组(80个克隆;插入约1.2 - 1.6 kb)与寡核苷酸混合物1-4杂交,而组II(12个克隆;插入物范围约1.2至1.4 kb)与寡核苷酸混合物杂交1-3。有几个组II的无性系比组I的插入体更大,但没有与寡核苷酸混合物4杂交。对人胎盘cDNA文库进行了450 bp的I组片段的筛选,也分离出I组和II组克隆。 Representative clones from group I (pASM-1) and group II (pASM-2) were sequenced. pASM-1 contained a 1879 bp insert which was colinear with 96 microsequenced amino acids, while the pASM-2 1382 bp insert was colinear with 78 microsequenced residues. Notably, pASM-2 did not have an internal 172 bp sequence encoding 57 amino acid residues, but had instead an in-frame 40 bp sequence encoding 13 amino acids which was not present in pASM-1. These findings demonstrate the presence of two distinct acid sphingomyelinase transcripts in human fibroblasts and placenta and suggest the occurrence of alternative processing of the mRNA encoding this lysosomal hydrolase.

引用本文的药物库数据

多肽
的名字 UniProt ID
鞘磷脂磷酸二酯酶 P17405 细节