Cloning of cDNA encoding the pre-beta subunit of mouse thyrotropin.
Cloning of cDNA encoding the pre-beta subunit of mouse thyrotropin.
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编码小鼠促甲状腺素前β亚基的cDNA的克隆。
DOI：
10.1073/pnas.80.8.2122
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发表时间：
1983
影响因子：
11.1
通讯作者：
Kourides,IA
中科院分区：
文献类型：
--
作者：
Gurr,JA;Catterall,JF;Kourides,IA
从小鼠促甲状腺肿瘤中提取聚腺苷酸（poly（A）+）mRNA，经蔗糖梯度纯化后，用聚腺苷酸（poly（dC））插入质粒pBR 322的Pst Ⅰ位点，合成双链cDNA。聚（dG）均聚物延伸，并在大肠杆菌RRI中克隆。通过杂交选择mRNA的无细胞翻译和与TSH β亚基特异性抗体的免疫沉淀，鉴定了含有编码促甲状腺激素（TSH）β亚基的cDNA序列的质粒。一个cDNA的核苷酸序列的测定，595个碱基对的长度，使我们能够推导出小鼠TSH β亚基的整个氨基酸序列。前β亚基含有20个氨基酸的氨基末端信号序列，随后是118个氨基酸的成熟TSH β亚基。小鼠TSH β亚基与人、猪和牛的亚基之间的氨基酸序列有85-90%的同源性;然而，与牛或人和猪亚基相比，小鼠亚基在其羧基末端分别含有额外的5或6个氨基酸。通过与标记的TSH β亚基cDNA杂交，估计小鼠促甲状腺肿瘤TSH β亚基mRNA的长度为750个核苷酸。
Double-stranded cDNA was synthesized from sucrose gradient-purified poly(A)+ mRNA from a mouse thyrotropic tumor, inserted into the Pst I site of plasmid pBR322 by using poly(dC) . poly(dG) homopolymeric extensions, and cloned in Escherichia coli RRI. Plasmids containing cDNA sequences coding for the beta subunit of thyrotropin (TSH) were identified by cell-free translation of hybrid-selected mRNA and immunoprecipitation with specific antibody to TSH beta subunit. Determination of the nucleotide sequence of one cDNA, 595 base pairs in length, allowed us to deduce the entire amino acid sequence of the mouse TSH beta subunit. The pre-beta subunit contains a 20-amino acid amino-terminal signal sequence followed by a 118-amino acid mature TSH beta subunit. There is 85-90% homology in amino acid sequence between mouse TSH beta subunit and subunits from man, pig, and cow; however, the mouse subunit contains an additional 5 or 6 amino acids at its carboxyl terminus compared to the bovine or human and pig subunits, respectively. TSH beta-subunit mRNA from mouse thyrotropic tumor was estimated to be 750 nucleotides in length by hybridization with labeled TSH beta-subunit cDNA.