Derynck R, Leung D W, Gray P W, Goeddel D V
Nucleic Acids Res. 1982 Jun 25;10(12):3605-15. doi: 10.1093/nar/10.12.3605.
Polyadenylated RNA from human peripheral blood lymphocytes and spleen cells, treated with different inducers for IFN-gamma production, was fractionated on denaturing sucrose gradients. Two IFN-gamma mRNA peaks at 12S and 16S were consistently observed. Nucleotide sequence analysis of cDNA clones showed that the 12S IFN-gamma mRNA from the different sources is identical to the gel fractionated 18S IFN-gamma mRNA which gave rise to the IFN-gamma cDNA clone p69 (1). Nucleotide sequence analysis of several IFN-gamma cDNA clones showed the presence of a CGA (Arg) codon at position 140 of mature IFN-gamma in contrast with the CAA (Gln) codon, which is found in p69 (1). Specifically primed cDNA extension on total induced polyadenylated RNA revealed the presence of a single mRNA species having a 5' untranslated region of 125-130 nucleotides. The nucleotide sequence of this region has been obtained. These data suggest that a single human IFN-gamma gene, which has very little polymorphism, gives rise to a single size class of mRNA.
用不同的γ干扰素产生诱导剂处理的人外周血淋巴细胞和脾细胞的聚腺苷酸化RNA,在变性蔗糖梯度上进行分级分离。始终观察到在12S和16S处有两个γ干扰素mRNA峰。cDNA克隆的核苷酸序列分析表明,来自不同来源的12Sγ干扰素mRNA与凝胶分级分离的18Sγ干扰素mRNA相同,后者产生了γ干扰素cDNA克隆p69(1)。几个γ干扰素cDNA克隆的核苷酸序列分析表明,与p69(1)中发现的CAA(谷氨酰胺)密码子相比,成熟γ干扰素第140位存在CGA(精氨酸)密码子。在总的诱导聚腺苷酸化RNA上进行特异性引物cDNA延伸,揭示存在一种单一的mRNA种类,其5'非翻译区为125-130个核苷酸。已获得该区域的核苷酸序列。这些数据表明,一个几乎没有多态性的单一人类γ干扰素基因产生单一大小类别的mRNA。