Kamb A, Weir M, Rudy B, Varmus H, Kenyon C
Department of Biochemistry and Biophysics, University of California, San Francisco 94143.
Proc Natl Acad Sci U S A. 1989 Jun;86(12):4372-6. doi: 10.1073/pnas.86.12.4372.
The study of gene family members has been aided by the isolation of related genes on the basis of DNA homology. We have adapted the polymerase chain reaction to screen animal genomes very rapidly and reliably for likely gene family members. Using conserved amino acid sequences to design degenerate oligonucleotide primers, we have shown that the genome of the nematode Caenorhabditis elegans contains sequences homologous to many Drosophila genes involved in pattern formation, including the segment polarity gene wingless (vertebrate int-1), and homeobox sequences characteristic of the Antennapedia, engrailed, and paired families. In addition, we have used this method to show that C. elegans contains at least five different sequences homologous to genes in the tyrosine kinase family. Lastly, we have isolated six potassium channel sequences from humans, a result that validates the utility of the method with large genomes and suggests that human potassium channel gene diversity may be extensive.
基于DNA同源性分离相关基因有助于对基因家族成员进行研究。我们采用聚合酶链反应,能够非常快速且可靠地筛选动物基因组,以寻找可能的基因家族成员。利用保守氨基酸序列设计简并寡核苷酸引物,我们发现线虫秀丽隐杆线虫的基因组包含与许多参与模式形成的果蝇基因同源的序列,包括节段极性基因无翅基因(脊椎动物int-1),以及触角足、成对控制和engrailed家族特有的同源异型框序列。此外,我们利用该方法表明秀丽隐杆线虫含有至少五个与酪氨酸激酶家族基因同源的不同序列。最后,我们从人类中分离出六个钾通道序列,这一结果证实了该方法在大型基因组研究中的实用性,并表明人类钾通道基因的多样性可能很广泛。