MOLECULAR GENETIC MANIPULATION OF THE DIATOM THALASSIOSIRA PSEUDONANA (BACILLARIOPHYCEAE)
作者： Nicole Poulsen , Patrick M. Chesley , Nils Kröger
DOI: 10.1111/J.1529-8817.2006.00269.X
关键词: Functional genomics 、 Transformation (genetics) 、 Biology 、 Computational biology 、 Botany 、 Transformation efficiency 、 Gene 、 Nourseothricin 、 Expression vector 、 Thalassiosira pseudonana 、 Marker gene
摘要: Here, we describe the first system for genetic transformation of Thalassiosira pseudonana (Hustedt) Hasle et Heimdal, only diatom which a complete genome sequence is presently available. This method based on microparticle bombardment followed by selection transformants using antibiotic nourseothricin. It exhibits highest efficiency compared with systems other species. To achieve high efficiency, it important to allow recovery bombarded T. cells in non-selective suspension culture before spreading nourseothricin containing agar plates. demonstrated that readily susceptible co-transformation allowing simultaneous introduction gene together marker gene. Both introduced genes are stably inherited even absence pressure. We have developed two pseudonana-specific expression vectors can drive constitutive (vector pTpfcp) and inducible pTpNR) genes. In combination available data expected provide powerful tool functional genomics diatoms.
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