转酿酒酵母海藻糖合成酶基因(TPS1)玉米植株的获得

TRANSFORMATION OF MAIZE WITH TREHALOSE SYNTHASE GENE(TPS1) CLONING FROM Saccharomyces cerevisiae

PCR克隆测序发现,酿酒酵母AS.1416菌株的海藻糖合成酶基因TPS1与原报道序列发生了11处单碱基突变,其中10个突变发生在编码区域,但9个是同义突变,不影响编码蛋白的氨基酸序列。只有1135位的G变为A,使编码蛋白的第355个甘氨酸变成了天冬酰胺。用单子叶植物逆境诱导启动子mwcs120启动TPS1基因构建表达载体,基因枪法转化玉米胚性愈伤组织,PCR检测获得阳性植株,平均达到0.56%的转化率。

Trehalose synthase gene （TPS1） was cloned from strain AS. 1416 of Saccharomyces cerevisiae by specific PCR. Sequence analysis showed that 11 single nucleotide mutations exist compared with the previously reported sequence. 10 of the mutations were located at the encoding domain, while 9 of them were synonymous mutations, which had no influence to the amino acid sequence of the encoding protein. Only the mutation from G to A at site 1 135 changed glycin of the 355th amino acid to asparagine. Expression vector with TPS 1 gene promoted by stress inducible promoter （ mwcs 120） of monocotyledon was used to transfer embyronic calli of maize by microprojectile bombardment. Positive plants were assayed by PCR amplification. The average transformation rate was 0.56 % .