title: [Establishment of BAC mediated transgenic mice containing 97 kb beta-globin gene cluster].
abstract: OBJECTIVE
  To delete IL-11 receptor alpha chain gene from the Bacterial Artificial Chromosome (BAC) chimeric DNA by RecA protein mediated homologous recombination method and establish the transgenic mice model containing whole beta-globin gene cluster.
  
  
  METHODS
  Two 500 bp homologous sequences (A and B) located at the upstream and downstream of IL-11 receptor alpha chain gene respectively were cloned into the Hind III and Xba I sites of pBV vector, then the 1 kb A + B fragment was recovered from the building vector and inserted into the Sal I site of the shuttle vector pSV-RecA. After transforming the shuttle vector into the competent DH10B E. Coli containing BAC DNA, the IL-11 receptor alpha chain gene was finally deleted from the BAC DNA through chloramphenicol positive selection and fusaic acid negative selection. The new BAC clone was characterized by Pulse Field Gel Electrophoresis (PFGE). Then, we microinjected the linearized and purified BAC DNA into the mouse fertilized eggs and prepared the transgenic mice.
  
  
  RESULTS
  By RecA protein mediated homologous recombination method, we deleted the IL-11 receptor alpha chain gene from the BAC DNA containing the complete beta-globin Gene Cluster and established 3 respective transgenic mice lines.
  
  
  CONCLUSIONS
  Human beta-globin gene cluster in the transgenic mice mediated by new BAC expresses in a correct mode and level as compared with previous transgenic mice.
authors.affiliation: National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Shen
authors.forename: Yue
authors.initials: Y
authors.lastname: Huang
authors.forename: Yi
authors.initials: Y
authors.lastname: Tang
authors.forename: De-pei
authors.initials: DP
authors.lastname: Liu
authors.forename: Guang
authors.initials: G
authors.lastname: Liu
authors.forename: Min
authors.initials: M
authors.lastname: Wu
authors.forename: Chih-chuan
authors.initials: CC
authors.lastname: Liang
mesh_terms: D000818:Animals; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D004247:DNA; D005914:Globins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D017370:Interleukin-11 / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D023421:Models, Animal; D005810:Multigene Family / Q000235:genetics*; D018123:Receptors, Interleukin / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014158:Transcription, Genetic
chemical_list: D017370:Interleukin-11; D018123:Receptors, Interleukin; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D005914:Globins; D004247:DNA