利用重叠PCR技术对OsAMT1.2进行定点突变

Point Mutant of OsAMT1.2 by Using Gene Splicing by Overlap Extension

铵是植物吸收利用的主要氮源之一,而铵转运蛋白对铵的吸收、转运和代谢方面有重要的作用。为了深入研究水稻铵转运蛋白的结构功能特性,利用重叠PCR技术对OsAMT1.2蛋白质序列中保守的253 aa和257 aa的苯丙氨酸(F)和丝氨酸(S)分别突变为丙氨酸,成功获得了相应的2个点突变基因。因此,重叠PCR技术是一种简单有效的进行体外基因重组和突变的方法,对于研究高等植物功能基因的结构功能特性具有重要的意义。

Ammonium is one of the important inorganic nitrogen in plant, and ammonium transporters are essential for ammonium uptake, transport, and metabolism. To further study the functional structure and characteristic for ammonium transporters in rice, highly conserved phenylalanine and serine residues, F253 and S257 of OsAMT1.2 were replaced by alanine respectively using gent splicing by overlap extension （gene SOEing） method. Results indicated that anticipant mutant genes were obtained successfully. Therefore, the technique of gene SOEing is a simple and effective method for gene recombination and mutation in Vitro, which is significant to study structurally and functional characteristic of gene in high plants.