Johanningmeier U, Hallick R B
Ruhr-University, Department Biochemistry of Plants, Bochum, Federal Republic of Germany.
Curr Genet. 1987;12(6):465-70. doi: 10.1007/BF00434825.
The psbA gene coding for the herbicide binding QB protein of photosystem II has been sequenced previously (Karabin et al. 1984). A herbicide resistant mutant of Euglena, Euglena gracilis ZR, was studied by sequencing part of the psbA gene and its corresponding mRNA. Sequencing reactions were done by annealing a psbA specific, end-labeled DNA-oligomer to total chloroplast DNA or RNA and extending this primer with reverse transcriptase in the presence of the four dideoxynucleotides. An amino acid substitution from serine to alanine at position 265 was detected. All known herbicide resistant higher plants sequenced to date and the Chlamydomonas mutant DCMU-4 show a change at exactly this same position, but the substitution in higher plants is from serine to glycine.
编码光系统II除草剂结合蛋白QB的psbA基因先前已被测序(卡拉宾等人,1984年)。通过对psbA基因的部分及其相应mRNA进行测序,研究了一种除草剂抗性突变体——纤细裸藻ZR。测序反应是通过将一个psbA特异性的、末端标记的DNA寡聚物与总叶绿体DNA或RNA退火,并在四种双脱氧核苷酸存在的情况下用逆转录酶延伸该引物来进行的。在第265位检测到从丝氨酸到丙氨酸的氨基酸替换。迄今为止测序的所有已知除草剂抗性高等植物以及衣藻突变体DCMU - 4在完全相同的位置都出现了变化,但高等植物中的替换是从丝氨酸到甘氨酸。