Gope M L, Keinänen R A, Kristo P A, Conneely O M, Beattie W G, Zarucki-Schulz T, O'Malley B W, Kulomaa M S
Nucleic Acids Res. 1987 Apr 24;15(8):3595-606. doi: 10.1093/nar/15.8.3595.
A cDNA for chicken avidin was identified in a chicken oviduct cDNA library by screening with antibodies and synthetic oligodeoxyribonucleotides. Four recombinant clones were characterized and each contained the sequence of the oligonucleotide probes used in screening. They were capable also of expressing an antigen recognizable by a polyclonal or a mixture of monoclonal antibodies raised against avidin. The longest clone, lambda cAV4, contained the entire coding sequence of avidin along with a signal peptide of 24 amino acids. An avidin mRNA, approximately 700 nucleotides in length, was induced by a single injection of progesterone over a period of twenty four hours. The avidin mRNA was distributed in a tissue-specific manner, since detectable concentration of the mRNA appeared only in the oviduct after stimulation with progesterone alone or with a combination of progesterone and estrogen. No avidin mRNA was detected in the liver or kidney under these conditions. Preliminary results on the genomic complexity of avidin suggest a single copy gene. Isolation of the natural gene for avidin and studies on its regulation now can be initiated using the cDNA probe.
通过用抗体和合成寡脱氧核糖核苷酸筛选，在鸡输卵管cDNA文库中鉴定出鸡抗生物素蛋白的cDNA。对四个重组克隆进行了表征，每个克隆都包含筛选中使用的寡核苷酸探针序列。它们还能够表达一种抗原，该抗原可被针对抗生物素蛋白产生的多克隆抗体或单克隆抗体混合物识别。最长的克隆λcAV4包含抗生物素蛋白的完整编码序列以及一个24个氨基酸的信号肽。单次注射孕酮在24小时内可诱导出长度约为700个核苷酸的抗生物素蛋白mRNA。抗生物素蛋白mRNA以组织特异性方式分布，因为仅在单独用孕酮或孕酮与雌激素联合刺激后，输卵管中才出现可检测浓度的mRNA。在这些条件下，肝脏或肾脏中未检测到抗生物素蛋白mRNA。抗生物素蛋白基因组复杂性的初步结果表明是单拷贝基因。现在可以使用cDNA探针开始抗生物素蛋白天然基因的分离及其调控研究。
