Construction of wild-type and mutant SPAST vectors for the study of molecular mechanism of hereditary spastic paraplegia.
- Author:
Ya-ping YAN
1
;
Jia-li PU
;
Bao-rong ZHANG
;
Guo-hua ZHAO
Author Information
- Publication Type:Journal Article
- MeSH: Adenosine Triphosphatases; genetics; metabolism; Animals; Base Sequence; Cell Line; Genetic Vectors; genetics; Humans; Mitochondria; genetics; metabolism; Mutation; Spastic Paraplegia, Hereditary; genetics; metabolism; Spastin
- From: Chinese Journal of Medical Genetics 2013;30(1):9-12
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct wild-type and mutant pEGFP SPAST vectors and to explore the molecular mechanism of hereditary spastic paraplegia.
METHODSMutant 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.
RESULTSWild-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.
CONCLUSIONWild-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.
