Chebath J, Tosta Z T, Chabaud O, Perricaudet M
Mol Biol Rep. 1982 Apr 16;8(3):149-56. doi: 10.1007/BF00777242.
This paper describes the cloning in E. coli of several DNA copies of the 3' portion of ovine thyroglobulin mRNA. The cDNA clones were identified by in situ cloning hybridization to 32P-labelled thyroglobulin cDNA and by positive hybridization-translation assays. The longest thyroglobulin cDNA fragment cloned (psTg21A, 1670 bp), was shown by S1 nuclease mapping to be an unaltered copy of the thyroglobulin mRNA. psTg 21A overlapped with another cDNA fragment (psTg 11, 330 bp) containing the poly(dA)-tail and corresponding therefore to the 3' end of the mRNA. When aligned together the two clones represent more than 20% of the thyroglobulin mRNA length. Another cDNA fragment (psTg 15, 350 bp) was identified as an internal portion of the thyroglobulin mRNA sequence.
本文描述了绵羊甲状腺球蛋白mRNA 3′端几个DNA拷贝在大肠杆菌中的克隆。通过与32P标记的甲状腺球蛋白cDNA进行原位克隆杂交以及阳性杂交翻译试验,鉴定出了这些cDNA克隆。通过S1核酸酶图谱分析表明,克隆得到的最长甲状腺球蛋白cDNA片段(psTg21A,1670 bp)是甲状腺球蛋白mRNA的未改变拷贝。psTg 21A与另一个含有聚(dA)尾且因此对应于mRNA 3′端的cDNA片段(psTg 11,330 bp)重叠。当这两个克隆排列在一起时,它们代表了甲状腺球蛋白mRNA长度的20%以上。另一个cDNA片段(psTg 15,350 bp)被鉴定为甲状腺球蛋白mRNA序列的内部部分。