Magnusson R P, Gestautas J, Seto P, Taurog A, Rapoport B
FEBS Lett. 1986 Nov 24;208(2):391-6. doi: 10.1016/0014-5793(86)81055-9.
We undertook the molecular cloning of porcine thyroid peroxidase (TPO). Four oligonucleotide probes were synthesized on the basis of amino acid sequences of 3 tryptic peptides from highly purified porcine TPO. These probes were used to screen a pig thyroid cDNA library. Seven of 16 selected clones (0.45-1.15 kb in size) reacted with all 4 probes. Nucleotide sequencing of the 1.15 kb at the 3'-end of the structural gene revealed the complementary sequence to all 4 probes as well as the nucleotides coding for the entire length of the 3 tryptic peptides. There is an open reading frame of 332 amino acid residues. On Northern blot analysis this gene codes for an mRNA species of 2.85 kb, corresponding to the anticipated size of the mRNA for the intact TPO molecule. We have therefore cloned and characterized a cDNA clone coding for approx. 36% of porcine thyroid peroxidase.
我们进行了猪甲状腺过氧化物酶（TPO）的分子克隆。根据高度纯化的猪TPO的3个胰蛋白酶肽段的氨基酸序列合成了4种寡核苷酸探针。这些探针用于筛选猪甲状腺cDNA文库。16个选定克隆中的7个（大小为0.45 - 1.15 kb）与所有4种探针发生反应。结构基因3'端1.15 kb的核苷酸测序揭示了与所有4种探针互补的序列以及编码3个胰蛋白酶肽段全长的核苷酸。存在一个332个氨基酸残基的开放阅读框。Northern印迹分析表明，该基因编码一种2.85 kb的mRNA，与完整TPO分子的预期mRNA大小相对应。因此，我们克隆并鉴定了一个编码约36%猪甲状腺过氧化物酶的cDNA克隆。
