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Plastid engineering of a marine alga, Nannochloropsis gaditana, for co-expression of two recombinant peptides

J Phycol. 2020-11; 
Yulin Cui, Kang Wang, Wenxin Xu, Yinchu Wang, Zhengquan Gao, Hongli Cui, Chunxiao Meng, Song Qin
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GenParts™ DNA Fragments … All the PCR products were electrophoresed on 0.8% agarose gel and the target bands were recovered using Quick Clean II gel extraction kit (GenScript, China). The purified fragments were sequenced by Ruibo Company. Plasmid construction … Get A Quote

摘要

The purpose of this study was to establish a plastid transformation system for expressing recombinant proteins in Nannochloropsis gaditana. On the basis of the sequenced plastid genome, the homologous flanking region, 16S-trnI/trnA-23S, and the endogenous regulatory fragments containing the psbA promoter, rbcL promoter, rbcL terminator, and psbA terminator were amplified from N. gaditana as elements of a plastid transformation vector. Then, the herbicide resistant gene (bar) was used as a selectable marker, regulated by the psbA promoter and rbcL terminator. Finally, two codon-optimized antimicrobial peptide-coding genes linked by endogenous ribosome binding site (RBS) in a polycistron were inserted into the co... More

关键词

Nannochloropsis gaditana, 16S-trnI/trnA-23S region, antimicrobial peptide, microparticle bombardment, plastid transformation, ribosome binding site