高羊茅CBF转录激活因子基因片段的克隆

MOLECULAR CLONING OF A GENE FRAGMENT ENCODING CBF TRANSCRIPTIONAL ACTIVATOR FROM TALL FESCUE

根据拟南芥CBF基因序列的保守区设计 1对特异引物 ,采用PCR方法从高羊茅基因组中扩增出 1个DNA片段并克隆到pUCm T载体中。经序列测定和分析 ,该片段长 60 8bp ,与 3个拟南芥CBF基因的核苷酸及其推导氨基酸序列分别具有 81～ 84%和 75～ 79%的同源性 ,而且推导氨基酸序列中包含有同源性更高的AP2DNA结合域以及CBF蛋白的两段特征序列PKK RPAGRxKFxETRHP和DSAWR。这些结果表明 。

A DNA fragment was amplified from genome of tall fescue by polymerase chain reaction using a pair of primers designed from the conserved sequences of three Arabidopsis CBF genes, and cloned into the vector pUCm T. Sequence measurement and analysis showed that the nucleotide and deduced amino acid sequences of the cloned 608 bp fragment were 81%～84% and 75%～79% identical to the three Arabidopsis CBF genes respectively. Moreover, there were an AP2 DNA binding domain of higher identity and two signature sequences of CBF proteins, PKK/RPAGRxKFxETRHP and DSAWR, in the deduced amino acid sequence. These results indicated that a tall fescue CBF gene fragment was obtained in this study.