title: [Construction and tumorigenic study on a novel fusion gene AML1-MTG16].
abstract: OBJECTIVE
  To test whether splicing overlapping extension(SOE) method can be a tool for obtaining rare fusion gene's transcripts and to study the tumorigenic capacity of a novel fusion gene AML1-MTG16.
  
  
  METHODS
  SOE method was used to obtain AML1-   MTG16 fusion gene's transcripts. MTG16, AML1-MTG16 and AML1-MTG16 without III,VI conserved domains of MTG16 segment were inserted into pEGFP-   C1,pDsRed-N1 vector respectively,then transfected NIH3T3 cell line by lipofection. Forty-eight hours later, the transfected cells were examined by laser-scanning confocal microscopy. Stable transfected cells were obtained by G418 500ug/ul selection for one month. Growth curve, soft agar colonies formation  tumorigenesis in nude mice were done to compare the difference between stable transfected cells.
  
  
  RESULTS
  Recombined AML1-MTG16 by SOE contained its CDS. NIH3T3 expressing AML1-MTG16 had a faster proliferation in medium, colony growth in soft agar. AML1-MTG16 expression cells also induced tumors formation following injection into nude mouse. MTG16,AML1-MTG16 and AML1-MTG16 without III,VI conserved domains of MTG16 were colocalized in the nucleus of cotransfected NIH3T3 cells under the examination of laser-scanning confocal microscope.
  
  
  CONCLUSIONS
  SOE is an effective method to get rare fusion gene's transcripts. AML1-MTG16 plays an important role in leukemogenesis. MTG16 may also have a carcinogenic property within the AML1-MTG16 fusion gene. Carcinogenic property of AML1-MTG16 is restricted to its localization in the nuclear matrix. N terminal of MTG16 may play an important part in the carcinogenic activity of AML1-MTG16.
authors.affiliation: Shanghai Institute of Hematology, Ruijin Hospital of Shanghai Second Medical University, Shanghai,200025 P.R. China. zhugangw@shsmu.edu.cn
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.forename: Shunyuan
authors.initials: S
authors.lastname: Lu
authors.forename: Hui
authors.initials: H
authors.lastname: Kong
authors.forename: Long
authors.initials: L
authors.lastname: Wang
authors.forename: Wentao
authors.initials: W
authors.lastname: Yuan
authors.forename: Yueping
authors.initials: Y
authors.lastname: Sun
authors.forename: Yue'e
authors.initials: Y
authors.lastname: Jing
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Lu
authors.forename: Zhenyi
authors.initials: Z
authors.lastname: Wang
authors.forename: Zhugang
authors.initials: Z
authors.lastname: Wang
mesh_terms: D016475:3T3 Cells / Q000637:transplantation; D000818:Animals; D002455:Cell Division / Q000235:genetics; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D017690:Cell Transplantation; D050676:Core Binding Factor Alpha 2 Subunit; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D018613:Microscopy, Confocal; D009374:Neoplasms, Experimental / Q000235:genetics / Q000473:pathology; D015514:Oncogene Proteins, Fusion / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics*; D014162:Transfection
chemical_list: C112643:AML1-MTG16 protein, human; D050676:Core Binding Factor Alpha 2 Subunit; D008164:Luminescent Proteins; D015514:Oncogene Proteins, Fusion; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D049452:Green Fluorescent Proteins