Delic-Attree I, Toussaint B, Vignais P M
CEA, Laboratoire de Biochimie Microbienne, CNRS URA 1130 alliée à l'INSERM, DBMS Centre d'Etudes Nucléaires, Grenoble, France.
Gene. 1995 Feb 27;154(1):61-4. doi: 10.1016/0378-1119(94)00875-s.
Histone-like proteins, such as HU and the integration host factor (IHF), are small, dimeric, DNA-bending proteins which play a role in maintaining constrained DNA structures and hence in regulating gene expression. Two different strategies were used to isolate the genes coding for Pseudomonas aeruginosa (Pa) HU and IHF, two proteins that we have previously isolated from a mucoid strain. By use of a PCR-based technique with oligodeoxyribonucleotides (oligos) designed from the N-terminal amino acid (aa) sequences of HU and the beta-subunit of IHF, and Southern blot analyses, hupB and himD, encoding HU and IHF beta, respectively, have been cloned. The himA gene of Pa, encoding the alpha-subunit of IHF, was isolated using himA of Escherichia coli (Ec) as a probe in Southern blot analyses. The deduced hupB product (90 aa, 9 kDa) is 79% identical to HU beta and 61% to HU alpha of Ec. The predicted products of himA (100 aa, 11.5 kDa) and of himD (94 aa, 10.6 kDa) share 77 and 70% identity with IHF alpha and IHF beta of Ec, respectively. The promoter region of himD contains an IHF consensus sequence, as is the case for Ec himD.
类组蛋白，如HU和整合宿主因子（IHF），是小型二聚体DNA弯曲蛋白，在维持受限的DNA结构以及调节基因表达中发挥作用。我们采用了两种不同策略来分离编码铜绿假单胞菌（Pa）HU和IHF的基因，这两种蛋白我们之前已从一株黏液型菌株中分离出来。通过使用基于聚合酶链反应（PCR）的技术，利用根据HU的N端氨基酸（aa）序列和IHF的β亚基设计的寡脱氧核糖核苷酸（oligos），以及Southern印迹分析，分别克隆了编码HU的hupB和编码IHFβ的himD。在Southern印迹分析中，以大肠杆菌（Ec）的himA作为探针，分离出了编码Pa的IHFα亚基的himA基因。推导的hupB产物（90个氨基酸，9 kDa）与Ec的HUβ有79%的同一性，与Ec的HUα有61%的同一性。预测的himA产物（100个氨基酸，11.5 kDa）和himD产物（94个氨基酸，10.6 kDa）分别与Ec的IHFα和IHFβ有77%和70%的同一性。himD的启动子区域含有一个IHF共有序列，Ec的himD也是如此。
