›› 2012, Vol. 39 ›› Issue (1): 23-27.

• 生物技术 • Previous Articles     Next Articles

Aspergillus usamii Xylanase Secrete Expression in Mammalian Cells

ZHANG Mao1, ZOU Xian2, LIN Chun2, LIU De-wu2, WU Zhen-fang2, CAI Geng-yuan1   

  1. 1. State Key Laboratory of Animal Breeding, Guangdong Public Laboratory of Animal Breeding and Nutrition, Institute of Animal Science, Guangdong Academy of Agricultural Sciences,Guangzhou 510640, China;2. College of Animal Science, South China Agricultural University,Guangzhou 510642, China
  • Received:1900-01-01 Revised:2011-06-21 Online:2012-01-20 Published:2012-01-20

Abstract: The mature peptide coding sequence of xylanase gene xyn (555 bp) was amplified by RT-PCR from Aspergillus niger GIM 3.36 total RNA extracts. It was subcloned into different locations of the eukaryotic expressing plasmid vector pcDNA6/HisTM A. The recombinant plasmid pcDNA-spna and pcDNA-spnb was identified by PCR, enzyme digestion and DNA sequencing. The result showed that the recombinant plasmids were constructed correctly. Meanwhile, the PK15 cells were transfected with pcDNA-spna and pcDNA-spnb by cationic liposome, and the mRNA of the target gene was determined by RT-PCR. The maximum yield of the recombinant xylanase (transfected pcDNA-spna) in cell culture medium was 8.53 U/mL and higher 24% the recombinant xylanase 6.87 U/mL (transfected pcDNA-spnb). To achieve the purpose that microbiology secrete expression in mammalian cells and provided basis for using xyn gene to transgenic.

Key words: Aspergillus usamii; xylanase; mammalian cells; secrete expression

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