Shamblott M J, Chen T T
Center of Marine Biotechnology, University of Maryland, Baltimore 21202.
Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):8913-7. doi: 10.1073/pnas.89.19.8913.
An internal portion of insulin-like growth factor (IGF) amplified from the total cDNA of rainbow trout (Oncorhynchus mykiss) liver by a PCR was used to screen a rainbow trout liver cDNA library, and recombinant clones encoding two distinct IGFs were isolated. On the basis of a 98.7% nucleotide and 98.3% predicted amino acid identity to coho salmon IGF-I, one cDNA sequence was identified as rainbow trout preproinsulin-like growth factor I (rtIGF-I). The second cDNA sequence shared 46.1% and 43.3% identity with rtIGF-I at the nucleotide and predicted amino acid levels, respectively, and was identified as rainbow trout preproinsulin-like growth factor II (rtIGF-II). Predicted amino acid sequence comparisons of rtIGFs with those of human IGFs indicate that rtIGF-I is more similar to human IGF-I than to human IGF-II, and that rtIGF-II is more similar to human IGF-II than to human IGF-I. Southern blot analysis of rainbow trout genomic DNA probed with rtIGF-I and -II cDNA suggests that these two forms of IGF originate from separate genes. The presence of a teleost IGF-II suggests that the divergence of IGFs occurred early in vertebrate evolution.
通过聚合酶链反应(PCR)从虹鳟(Oncorhynchus mykiss)肝脏的总cDNA中扩增出的胰岛素样生长因子(IGF)内部片段,用于筛选虹鳟肝脏cDNA文库,并分离出编码两种不同IGF的重组克隆。基于与银大麻哈鱼IGF-I的98.7%核苷酸和98.3%预测氨基酸同一性,一个cDNA序列被鉴定为虹鳟前胰岛素样生长因子I(rtIGF-I)。第二个cDNA序列在核苷酸和预测氨基酸水平上分别与rtIGF-I具有46.1%和43.3%的同一性,并被鉴定为虹鳟前胰岛素样生长因子II(rtIGF-II)。rtIGF与人类IGF的预测氨基酸序列比较表明,rtIGF-I与人类IGF-I比与人类IGF-II更相似,并且rtIGF-II与人类IGF-II比与人类IGF-I更相似。用rtIGF-I和-II cDNA探测虹鳟基因组DNA的Southern印迹分析表明,这两种形式的IGF起源于不同的基因。硬骨鱼IGF-II的存在表明IGF的分化在脊椎动物进化早期就已发生。