title: [Construction of wild-type and mutant SPAST vectors for the study of molecular mechanism of hereditary spastic paraplegia].
abstract: OBJECTIVE
  To construct wild-type and mutant pEGFP SPAST vectors and to explore the molecular mechanism of hereditary spastic paraplegia.
  
  
  METHODS
  Mutant SPAST vector was constructed using overlap PCR method following construction of wild-type SPAST vector. Wild-type and mutant constructs were transfected to COS7 cells and subcellular localization of spastin was observed. Co-localizations of spastin and microtubule, spastin and mitochondria were viewed by immunofluorescence staining.
  
  
  RESULTS
  Wild-type spastin is localized in plasma, and mutant spastin did not change its cellular localization. Wild-type and mutant spastins did not co-localize with microtubules and mitochondria by immunofluorescence analysis.
  
  
  CONCLUSIONS
  Wild-type and mutant SPAST constructs were successfully generated. Mutant spastin did not change its localization in cells. Spastin does not co-localize with microtubules and mitochondria. This study may facilitate further studies on molecular mechanism of hereditary spastic paraplegia.
authors: Department of Neurology, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
authors: Ya-ping
authors: YP
authors: Yan
authors: Jia-li
authors: JL
authors: Pu
authors: Bao-rong
authors: BR
authors: Zhang
authors: Guo-hua
authors: GH
authors: Zhao
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D015419:Spastic Paraplegia, Hereditary / Q000235:genetics* / Q000378:metabolism; D000074181:Spastin
chemical_list: D000251:Adenosine Triphosphatases; D000074181:Spastin; C400283:SPAST protein, human