人类酸性鞘磷脂酶。隔离,全身的核苷酸序列和表达,或者拼接的互补。
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Schuchman呃,Suchi M,高桥T, Sandhoff K, Desnick RJ
人类酸性鞘磷脂酶。隔离,全身的核苷酸序列和表达,或者拼接的互补。
生物化学杂志。1991年5月5日,266 (13):8531 - 9。
- PubMed ID
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1840600 (在PubMed]
- 文摘
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两种类型的部分互补编码人类酸性鞘磷脂酶(EC 3.1.4.12;ASM)最近被隔绝成纤维细胞和胎盘cDNA库(Quintern l E。Schuchman,再见Levran, O。,Suchi。M。Ferlinz, K。Reinke, H。Sandhoff, K。,Desnick EMBO j . r . j . (1989) 2469 - 2473)。互补dna插入相同的序列与内部区域的异常;1型互补(代表大约90%的ASM的互补隔离)有172在坐标系碱基对(bp),取代的2型由40-bp互补序列在坐标系。北方杂交和核糖核酸酶保护研究表明1型和2成绩单都是大约2.5个碱基;因此,工作是针对孤立全身的人类胎盘类型1和2的互补筛选,睾丸,肝癌,视网膜互补脱氧核糖核酸数据库。 In addition to type 1 and 2 cDNAs, a new type of ASM cDNA (type 3), which did not contain the type 1- or 2-specific regions, was isolated and sequenced. The full-length type 1 and the reconstructed full-length type 2 and 3 cDNAs were transiently expressed in COS-1 cells. Only the full-length type 1 transcript encoded catalytically active human ASM, demonstrating its functional integrity. The 2347-bp full-length type 1 placental cDNA (pASM-1FL) had an 87-bp 5'-untranslated region, an 1890-bp open reading frame encoding 629 amino acids, and a 370-bp 3'-untranslated sequence. The predicted location of the signal peptide cleavage site was after alanine 46. Two base differences were identified in codons 322 and 506 and shown to be polymorphisms with the common alleles having frequencies of 0.6 and 0.7, respectively. To determine the genomic organization of the type 1, 2, and 3 sequences, a 1665-bp genomic region containing both the unique type 1 (172 bp) and type 2 (40 bp) sequences was amplified by the polymerase chain reaction and sequenced. The 172-bp sequence was exonic, flanked by 5'- and 3'-intronic sequences of 1052 and 229 bp, respectively. The 40-bp type 2 sequence was intronic, occurring at the 5' end of the 1052-bp intron due to the use of a cryptic 5' donor splice site, which deleted the entire 172-bp exon and both flanking intronic sequences. The type 3 cDNA resulted from an alternative splicing event, which excised the 172-bp exon. These studies demonstrate the occurrence of alternatively splicing of the ASM transcript, but the existence of only one functional mRNA.