Ullrich A, Berman C H, Dull T J, Gray A, Lee J M
EMBO J. 1984 Feb;3(2):361-4. doi: 10.1002/j.1460-2075.1984.tb01812.x.
A single synthetic oligonucleotide was employed as hybridization probe to detect and enable isolation of the human insulin-like growth factor I (IGF-I) gene from a human genomic DNA library. The synthetic oligonucleotide probe coded for the B-chain of IGF-I and was designed for expression in Escherichia coli. Despite numerous interspersed mismatches, the synthetic probe hybridized specifically with seven recombinant lambda phage containing almost the entire B-chain region of the human IGF-I gene. The usefulness of this approach was further demonstrated by the detection of lambda phage containing human preproinsulin, using A and B chain synthetic oligonucleotides, 90 and 63 nucleotides in length, as hybridization probes. The nucleotide sequence of the human IGF-I exon suggests that IGF-I is synthesized as a larger precursor molecule.
使用单个合成寡核苷酸作为杂交探针,从人基因组DNA文库中检测并分离人胰岛素样生长因子I(IGF-I)基因。该合成寡核苷酸探针编码IGF-I的B链,并设计用于在大肠杆菌中表达。尽管存在许多散布的错配,但合成探针与七个重组λ噬菌体特异性杂交,这些噬菌体包含几乎整个人IGF-I基因的B链区域。通过使用长度分别为90和63个核苷酸的A链和B链合成寡核苷酸作为杂交探针,检测含有人胰岛素原的λ噬菌体,进一步证明了该方法的实用性。人IGF-I外显子的核苷酸序列表明,IGF-I是作为一个更大的前体分子合成的。