塔乌库姆冰草肌动蛋白基因片段的克隆及序列分析

摘要

Degenerate primers were designed based on the conserved sequences of the actin gene according to other plants.Total RNA was extracted from the leaves of Agropyron cristatum cv.Tawukumu.Actin gene frag-ment was obtained by reverse transcription polymerase chain reaction (RT-PCR)and cloned into vector.The positive clone identified by PCR was sequenced.The sequencing result revealed that the Actin gene fragment from Agropyron cristatum cv.Tawukumu contains about 600 bp,encoding 1 99 amino acids.Homology compari-son with Actin gene sequences of other plants in the GenBank and with other actins showed that it shared over 81% nucleotide sequence homology and 88% amino acid sequence homology.%根据其他植物 Actin 基因的保守序列设计一对简引并性物,以塔乌库姆冰草叶片总 RNA 为模板,采用 RT-PCR 的方法扩增出 Actin 基因片段并连接到载体上,阳性克隆经 PCR 检测后进行测序。结果表明：该片段长约600 bp,编码199个氨基酸,所得序列与 GenBank 中注册的其他植物 Actin 基因序列同源性均在81%以上,与其他肌动蛋白的氨基酸序列同源性达88%。