Construction of middle fragment deletion mutant with improved gene splicing by overlap extension / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
;
(6): 52-55, 2004.
Article
in Chinese
| WPRIM
| ID: wpr-329400
ABSTRACT
<p><b>OBJECTIVE</b>To construct a phosphatidylinositol 4-kinase beta (PI4K-beta) mutant with the 325th to 373rd amino acid codons deleted, and try to develop a simple method for constructing middle fragment deletion mutant.</p><p><b>METHODS</b>In line with the mechanism of gene splicing by overlap extension(SOE), an additional PCR was used to get the PI4K-beta mutant in which the 325th to 373rd amino acid codons were deleted. Then the mutated gene was cloned into pCMV-Tag4A mammalian expression vector.</p><p><b>RESULTS</b>A mutant with the 325th to 373rd amino acid codons deleted was constructed successfully.</p><p><b>CONCLUSION</b>The improved SOE is a very effective and reliable method to construct middle fragment deletion mutant. It is worthy to be popularized.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Recombinant Proteins
/
Molecular Sequence Data
/
Base Sequence
/
Protein Engineering
/
Polymerase Chain Reaction
/
Mutagenesis, Site-Directed
/
Sequence Deletion
/
1-Phosphatidylinositol 4-Kinase
/
Genetic Vectors
/
Genetics
Language:
Chinese
Journal:
Chinese Journal of Medical Genetics
Year:
2004
Type:
Article
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