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Plastid Engineering of a Marine Alga, Nannochloropsis gaditana , for Co‐Expression of Two Recombinant Peptides
Author(s) -
Cui Yulin,
Wang Kang,
Xu Wenxin,
Wang Yinchu,
Gao Zhengquan,
Cui Hongli,
Meng Chunxiao,
Qin Song
Publication year - 2021
Publication title -
journal of phycology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.85
H-Index - 127
eISSN - 1529-8817
pISSN - 0022-3646
DOI - 10.1111/jpy.13099
Subject(s) - terminator (solar) , biology , plastid , recombinant dna , gene , transformation (genetics) , selectable marker , expression vector , microbiology and biotechnology , genetics , chloroplast , ionosphere , physics , astronomy
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 psb A promoter, rbc L promoter, rbc L terminator, and psb A 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 psb A promoter and rbc L terminator. Finally, two codon‐optimized antimicrobial peptide‐coding genes linked by endogenous ribosome binding site (RBS) in a polycistron were inserted into the constructed vector under the regulation of the rbc L promoter and psb A terminator. After microparticle bombardment, the positive clones were detected using polymerase chain reaction (PCR), and Southern and Western blotting were used to assess the co‐expression of the two antimicrobial peptides from the plastid. Nannochloropsis gaditana showed the potential to express recombinant proteins for biotechnological applications, for example, for the development of oral vaccines in aquaculture.