Keshet E, Rosner A, Bernstein Y, Gorecki M, Aviv H
Nucleic Acids Res. 1981 Jan 10;9(1):19-30. doi: 10.1093/nar/9.1.19.
A hybrid plasmid was constructed containing beta-lactamase gene of plasmid pBR322 and cloned coding sequences of bovine growth hormone (BGH). The constructed plasmid contains all DNA sequences required to encode BGH, and when used as a hybridization probe it detects one growth hormone gene in the bovine genome. The cloned DNA sequences are inserted into the beta-lactamase gene in the correct reading frame for BGH synthesis. The hybrid gene is expressed in bacteria and the product, a fused beta-lactamase-bovine growth hormone protein, is specifically immunoprecipitated with anti-serum to BGH. Unlike beta-lactamase, very little growth hormone containing sequences can be detected in the periplasmic space.
构建了一种杂种质粒,它含有质粒pBR322的β-内酰胺酶基因以及牛生长激素(BGH)的克隆编码序列。构建的质粒包含编码BGH所需的所有DNA序列,并且当用作杂交探针时,它能检测到牛基因组中的一个生长激素基因。克隆的DNA序列以正确的阅读框插入到β-内酰胺酶基因中以合成BGH。杂种基因在细菌中表达,产物即融合的β-内酰胺酶-牛生长激素蛋白,能被抗BGH血清特异性免疫沉淀。与β-内酰胺酶不同,在周质空间中只能检测到极少量含生长激素的序列。