Cellulase and Macerozyme-PEG-mediated Transformation of Moss Protoplasts

纤维素酶和离析酶-PEG 介导的苔藓原生质体转化

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作者:Rituraj Batth, Ilenuta Simina Cuciurean, Satish Kumar Kodiripaka, Sarah Skovlund Rothman, Caroline Greisen, Henrik Toft Simonsen

Abstract

This protocol describes the generation of protoplasts from protonemal tissue of the moss Physcomitrium patens (syn. Physcomitrella patens), using Cellulase ONOZUKA R10 and Macerozyme R10, followed by polyethylene glycol (PEG) mediated transformation. The protonemal tissue grown in liquid suspension was harvested and treated with enzyme cocktails mix of 1.5% Cellulase ONOZUKA R10 and 0.5% Macerozyme R10 to generate 1,8 million protoplasts within 3 h.

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