Journal of Capital Medical University ›› 2004, Vol. 25 ›› Issue (1): 35-39.

• 论著·基础研究 • Previous Articles     Next Articles

Construction and Identification of pPICZα-synBPIm600 Recombinant Expression Vector

Cong Min1, Jing Xuefang2, Liu Zhenlong3, Sun Mingjie3, An Yunqing3   

  1. 1. Liver Center, Beijing Friendship Hospital, Affiliate of Capital University of Medical Sciences;2. Docter of Immunology, Jilin University;3. Department of Immunology, Capital University of Medical Sciences
  • Received:2003-10-14 Revised:1900-01-01 Online:2004-01-15 Published:2004-01-15

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Abstract: In order to express rBPIm23 recombinant bactericidal protein in Pichia pa storis, synBPIm600 expression vectors were constructed. synBPI200 bp fragment was amplified by PCR from pUC18-synBPI414 vector, then the product was diges ted by XhoI and EcoRI to obtain synBPI185 bp fragment; PBV220-BPIm600 recombinant expressing vector was digested by EcoRI and SalI to obtain BPIm420 bp fragment; the above two fragments were linked and cloned in pPICZα e xpression vector to form intact pPICZα-synBPIm600 recombinant expression vector. After E.coli TOP 10 F' was transformated by pPICZα-synBPIm600 vector, positive cloning strains were selected by low salt LB medium with Zeocin and id entificated by PCR amplication, restriction analysis by XhoI/EcoRI/SalI and DNA sequencing. The results show that intact pPICZα-synBPIm600 recombinant expression vectors are constructed successfully.

Key words: bactericidal/permeability-incr easing protein(BPI), rBPIm23 recombinant bactericidal protein, ex pression vector, Pichia pastoris

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