Herbicide basta resistant gen bar as a new selective marker for genetic engineering of Phaeodactylum tricornutum
除草剂草丁膦抗性基因bar作为三角褐指藻基因工程选择标记的研究

Zeocin, a routine selective reagent in diatom genetic engineering, is not suitable for the construction of expression system of diatoms, due to its photosensitivity and mutagenicity. In order to find a suitable selective reagent for photobioreactor-based culture of transformed Phaeodactylum tricornutum. The sensitivity of P. tricornutum to five antibiotics (Chloramphenicol, Kanamycin, Penicillin, Streptomycin, and Hygromycin) and phosphinothricin (PPT) was tested. The results showed that P. tricornutum was sensitive to PPT, with a 72-h half death concentration of 22 mg/L. The expression vector pSVB, which contained PPT-resistant gene bar, was transformed into P. tricornutum cells by particle bombardment; and the resistant P. tricornutum was selected by PPT screening. It was perimentally confirmed that the transformed P. tricornutum was resistent to PPT. This research indicates that phosphinothrin is suitable for constructing a safe, efficient, low cost expression system of P. tricornutum.