Soma G I, Kitahara N, Nishizawa T, Nanami H, Kotake C, Okazaki H, Andoh T
Nucleic Acids Res. 1984 Nov 12;12(21):8029-41. doi: 10.1093/nar/12.21.8029.
We have isolated a cDNA clone encoding salmon proopiomelanocortin precursor. Polyadenylated RNA was isolated from pituitary neurointermediate lobes and used to construct a cDNA library. The library was screened with 17 mer of oligodeoxyribonucleotides specific for the hexapeptide sequence in salmon beta-endorphin I, Phe-Met-Lys-Pro-Tyr-Thr at positions 4-9 excluding the third nucleotide. One positive clone, pSSM17 containing an insert of 1303 base pairs (bp) was characterized. Sequence determination revealed that it possessed sequences covering the entire regions encoding ACTH and beta-lipotropin and that the mRNA had the same overall organization as those of other mammalian species, i.e., the following peptide hormones were arranged in order from 5' upstream, ACTH including alpha-melanotropin and corticotropin-like intermediate lobe peptide, beta-lipotropin including gamma-lipotropin, beta-melanotropin and beta-endorphin. Amino acid sequences for putative salmon ACTH, beta-, and gamma-lipotropin were predicted. Comparison of the salmon mRNA sequence with those of mammals showed that the regions of alpha- and beta-MSH are relatively homologous, but other regions are much less so, especially in the 3' nontranslated region where it is much longer and completely heterologous.
我们已分离出一个编码鲑鱼阿片促黑素皮质素原前体的cDNA克隆。从垂体神经中间叶分离出聚腺苷酸化RNA,并用于构建cDNA文库。用针对鲑鱼β-内啡肽I中六肽序列(4-9位的苯丙氨酸-甲硫氨酸-赖氨酸-脯氨酸-酪氨酸-苏氨酸,不包括第三个核苷酸)的17聚体寡脱氧核糖核苷酸筛选该文库。对一个阳性克隆pSSM17进行了鉴定,其插入片段为1303个碱基对(bp)。序列测定表明,它具有覆盖编码促肾上腺皮质激素(ACTH)和β-促脂素整个区域的序列,并且该mRNA的总体结构与其他哺乳动物物种的相同,即以下肽激素从5'上游开始依次排列:ACTH(包括α-促黑素和促肾上腺皮质激素样中间叶肽)、β-促脂素(包括γ-促脂素、β-促黑素和β-内啡肽)。预测了假定的鲑鱼ACTH、β-和γ-促脂素的氨基酸序列。将鲑鱼mRNA序列与哺乳动物的序列进行比较表明,α-和β-促黑素区域相对同源,但其他区域的同源性要低得多,尤其是在3'非翻译区,该区域长得多且完全异源。