Uhler M, Herbert E
J Biol Chem. 1983 Jan 10;258(1):257-61.
Polyadenylated RNA was isolated from a mouse pituitary tumor cell line (AtT-20/D16v) which synthesizes and secretes adrenocorticotropic hormone and beta-endorphin. The RNA was used to construct a cDNA library by a double linker technique and the library was screened for pro-opiomelanocortin (POMC) sequences. One recombinant plasmid, pMKSU16, contained a 923-base pair insert comprising the entire POMC-coding sequence as well as 98 bases of 5' noncoding and 5 bases of 3' noncoding sequence. The protein sequence predicted by the cDNA shows mouse POMC to consist of 235 amino acids with seven potential tryptic cleavage sites consisting of pairs of basic amino acid residues. Comparison with the previously published bovine POMC sequence suggests certain regions of POMC are highly conserved between the two species, particularly the regions corresponding to the alpha-, beta-, and gamma-melanocyte-stimulating hormones.
多聚腺苷酸化RNA是从小鼠垂体肿瘤细胞系(AtT - 20/D16v)中分离得到的,该细胞系能合成并分泌促肾上腺皮质激素和β-内啡肽。利用双链连接技术,用该RNA构建了一个cDNA文库,并筛选其中的阿黑皮素原(POMC)序列。一个重组质粒pMKSU16含有一个923个碱基对的插入片段,该片段包含整个POMC编码序列以及5'非编码区的98个碱基和3'非编码区的5个碱基。由该cDNA预测的蛋白质序列显示,小鼠POMC由235个氨基酸组成,有7个潜在的胰蛋白酶切割位点,这些位点由成对的碱性氨基酸残基组成。与先前发表的牛POMC序列进行比较表明,POMC的某些区域在这两个物种之间高度保守,特别是对应于α-、β-和γ-黑素细胞刺激素的区域。