Construction of Novel Chloroplast Expression Vector and Development of an Efficient Transformation System for the Diatom Phaeodactylum tricornutum

• DOI: 10.1007/s10126-014-9570-3
• Publication Date: 2014-04-25T02:25:12Z
• Authors: Wei-Hong Xie; Cong-Cong Zhu; Nai-Sheng Zhang; Da-Wei Li; Wei-Dong Yang; Jie-Sheng Liu; Ramalingam Sathis...; Hong-Ye Li
• Keywords: Phaeodactylum tricornutum; Heterologous expression; Chloramphenicol acetyltransferase; Translational efficiency

Plastids are ideal subcellular hosts for the expression of transgenes and have been successfully used production different biopolymers, therapeutic proteins industrial enzymes. Phaeodactylum tricornutum is a widely aquatic feed species. In this study, we focused on developing high-efficiency plastid system P. tricornutum. transformation vector, site selected integration was transcriptionally active intergenic region present between trnI trnA genes, located in IR (inverted repeat) regions genome. Initially, CAT reporter gene (encoding chloramphenicol acetyltransferase) integrated at The transformed microalgae conferred resistance to antibiotic chloramphenicol, which enabled growth selection media. Overall, efficiency found be approximately one transplastomic colony per 1,000 cells. Subsequently, heterologous cassette high-level target created cloned homologous recombination elements. A TA cloning strategy based designed XcmI-XcmI sites could conveniently clone gene. An eGFP (green fluorescent protein) test level system. relatively without codon optimisation stably determined account 0.12 % total soluble protein. Thus, study presents first convenient diatoms represents an interesting tool diatom plastids.