Elevated expression of human alpha-1 antitrypsin mediated by yeast intron in Pichia pastoris.
Intron-mediated enhancement has been documented in many cases to involve large positive effect on gene expression. To address this, human Alpha-1 antitrypsin (hAAT) gene was integrated into Pichia pastoris with and without a yeast intron generated from the final plasmid pBlu-exII-int-exIII and ligated into the EcoRI/BamHI multiple cloning site of the yeast shuttle vector pHIL-S1. The chimeric exon-intron complex in the middle of the naturally occurring hAAT exons II and III caused a 23-fold enhancement of hAAT expression in P. pastoris, measured through SDS-PAGE and immunoblot analyses.
Hasannia
S
S
Department of Clinical Biochemistry, Tarbiat Modarres University, Tehran, Iran.
Lotfi
A S
AS
Mahboudi
F
F
Rezaii
A
A
Rahbarizadeh
F
F
Mohsenifar
A
A
D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D015151:Immunoblotting; D007438:Introns / Q000235:genetics*; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D019076:Transgenes; D014361:Trypsin Inhibitors / Q000096:biosynthesis / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics*; D000515:alpha 1-Antitrypsin / Q000096:biosynthesis* / Q000235:genetics*
D011994:Recombinant Proteins; D014361:Trypsin Inhibitors; D000515:alpha 1-Antitrypsin