Guo W, Worley K, Adams V, Mason J, Sylvester-Jackson D, Zhang Y H, Towbin J A, Fogt D D, Madu S, Wheeler D A
Institute for Molecular Genetics, Baylor College of Medicine, Houston, Texas 77030.
Nat Genet. 1993 Aug;4(4):367-72. doi: 10.1038/ng0893-367.
Rapid genomic scanning methods are required to identify expressed sequences and we report an efficient, sensitive and specific approach which relies upon hybridization of an amplified, labeled cDNA library to digested cosmid DNA. We identified expressed sequences within a cosmid in the glycerol kinase (GK) "critical region" of Xp21 that had impressive similarity to prokaryotic GKs. We used this genomic sequence information to clone the human hepatic GK cDNA. Independent confirmation of the identity of this gene was obtained by functional complementation of GK deficient E. coli mutants with a construct containing the complete human X-linked GK coding sequence.
需要快速基因组扫描方法来鉴定表达序列，我们报告了一种高效、灵敏且特异的方法，该方法依赖于将扩增的、标记的cDNA文库与消化的黏粒DNA进行杂交。我们在Xp21的甘油激酶（GK）“关键区域”的一个黏粒中鉴定出表达序列，这些序列与原核GK有惊人的相似性。我们利用这些基因组序列信息克隆了人肝GK cDNA。通过用含有完整人类X连锁GK编码序列的构建体对GK缺陷型大肠杆菌突变体进行功能互补，独立证实了该基因的身份。
