Halford S E, Jordan S L, Kirkbride E A
Mol Gen Genet. 1985;200(1):169-75. doi: 10.1007/BF00383331.
The tac promoter was inserted into Tn21 upstream of the tnpR gene and the resultant plasmid was used to generate substantial amounts of resolvase. This protein was purified to homogeneity. The protein was characterized by amino acid sequence studies (which showed that an open-reading frame previously identified by DNA sequencing had been correctly assigned to the tnpR gene) and by molecular weight measurements (which demonstrated that the only active for of the protein in solution was dimeric). Pure Tn21 resolvase catalysed site-specific recombinations between directly repeated res sites from Tn21 or Tn1721 but not from Tn3 nor on inverted res sites from Tn21.
将 tac 启动子插入到 Tn21 的 tnpR 基因上游,所得质粒用于大量产生解离酶。该蛋白质被纯化至同质状态。通过氨基酸序列研究(表明先前通过 DNA 测序鉴定的开放阅读框已正确定位到 tnpR 基因)和分子量测量(证明该蛋白质在溶液中的唯一活性形式为二聚体)对该蛋白质进行了表征。纯 Tn21 解离酶催化 Tn21 或 Tn1721 中直接重复的 res 位点之间的位点特异性重组,但不催化 Tn3 的 res 位点之间或 Tn21 的反向 res 位点之间的重组。