STABLE PLASTID TRANSFORMATION IN PEG-TREATED PROTOPLASTS OF NICOTIANA-TABACUM

被引：100
|
作者：
GOLDS, T
MALIGA, P
KOOP, HU
机构：
[1] UNIV MUNICH,PLANT CELL BIOL & CELL CULTURE LAB,W-8000 MUNICH 2,GERMANY
[2] RUTGERS UNIV,WAKSMAN INST,PISCATAWAY,NJ 08855
来源：
BIO-TECHNOLOGY | 1993年 / 11卷 / 01期
关键词：
D O I：
10.1038/nbt0193-95
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
Plastid engineering currently relies on DNA delivery by the biolistic process. We report here stable plastid transformation in tobacco by an alternate direct transformation protocol that is based on polyethylene glycol (PEG) treatment of leaf protoplasts in the presence of the transforming DNA. Clones with transformed plastid genomes were selected by spectinomycin resistance encoded by a mutant 16S ribosomal RNA gene. Incorporation of the transforming DNA into the plastid genome was confirmed by two unselected markers, streptomycin resistance and a novel PstI site that flank the spectinomycin resistance mutation in plasmid pZS148. Our simple and inexpensive protocol eliminates the dependence on the particle gun for chloroplast transformation and should facilitate applications of plastome engineering in crops.
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页码：95 / 97
页数：3
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