Article

Journal of Applied Phycology

, Volume 25, Issue 1, pp 139-144

Expression of soybean Kunitz trypsin inhibitor gene SKTI in Dunaliella salina

  • Xiao-Jie ChaiAffiliated withKey Laboratory of Resource Recovery and Environment Restoration of Marine Biology, Dalian Ocean UniversityDepartment of Fisheries and Life Science, Dalian Ocean University Email author 
  • , Hui-Xia ChenAffiliated withKey Laboratory of Resource Recovery and Environment Restoration of Marine Biology, Dalian Ocean University
  • , Wen-Qi XuAffiliated withKey Laboratory of Resource Recovery and Environment Restoration of Marine Biology, Dalian Ocean University
  • , Ya-Wei XuAffiliated withJilin College of Agricultural Science and Technology

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Abstract

Recently, microalgae have attracted much attention as a new bioreactor system for producing high-value heterologous proteins. In this paper, we investigated the expression of soybean Kunitz trypsin inhibitor gene SKTI in the chlorophyte Dunaliella salina. Using D. salina genomic DNA as template, the entire coding region of SKTI was amplified by PCR as a 654-bp DNA fragment. It had 100 % identity with the published sequence of SKTI. The entire SKTI fragment was cloned into the expression vector pCAM2201 at a site just downstream of 35S promoter of to yield pCAMSKTI. D. salina cells transfected with pCAMSKTI by means of the lithium acetate/polyethylene glycol-mediated method expressed a protein of 20.1 kDa as detected by anti-SKTI antibody. In addition, SDS-PAGE analysis of the cell extract also revealed an intense protein band indicative of the recombinant SKTI. SKTI was therefore successfully expressed in D. salina, and the expression could be detected for at least 35 generations.

Keywords

Soybean Kunitz trypsin inhibitor gene SKTI Dunaliella salina LiAc/PEG-mediated method