Expression of Chinese shrimp Fenneropenaeus chinensis ALFFc in Pichia pastoris

doi:10.3969/j.issn.1000-1565.2018.01.010

Abstract

Abstract: Anti-lipopolysaccharide factor(ALF)is a low molecular mass protein with antimicrobial activities and plays an important role in immune response against microbial invasion.Thus, they are considered to be good substitutes for traditional antibiotics.In the present study, the prokaryotic recombinant expression vector “pET-DsbA-ALFFc” was used as the template of PCR, which was constructed in previous study.EcoRI and NotI restriction sites were added to 5’ and 3’ ends of the ALFFc.The ALFFc fragment was ligated to pPIC9K vector to construct a recombinant expression vector pPIC9K-ALFFc, which then was transformed into competent Pichia pastoris GS115.Recombinant ALFFc(rALFFc)was induced with methanol and purified by Ni-chelating affinity chromatography.SDS-PAGE and Western blot analysis indicated that the rALFFc was expressed successfully in P.pastoris.

Key words: Fenneropenaeus chinensis, anti-lipopolysaccharide factor(ALF), Pichia pastoris, recombinant expression

CLC Number:

TANG Ting, LU Meike, MA Xinjian, LI Ting, LIU Fengsong. Expression of Chinese shrimp Fenneropenaeus chinensis ALFFc in Pichia pastoris[J]. Journal of Hebei University (Natural Science Edition), 2018, 38(1): 59-64.

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