新疆小拟南芥ApCBF1基因的克隆及其过量表达转基因的研究

cDNA Cloning of ApCBF1 Gene and Over-expression of ApCBF1 in Arabidopsis pumila

利用同源克隆法,克隆了小拟南芥的CBF1基因的cDNA(FJ491244),比对分析结果表明,ApCBF1的cDNA序列的A252,在拟南芥的AtCBF1的cDNA是T252,但翻译的氨基酸均是Ala,表明ApCBF1和AtCBF1翻译的氨基酸序列完全一致。构建了过量表达载体:p35S:ApCBF1,通过花滴法转入小拟南芥中。RT-PCR检测结果表明,转基因T0代AtCBF1基因过量表达。

Transcription activator CBF is the critical regulator in plant cold domestication. The cDNA sequence of CBF1 in Arabidopsis pumila was obtained through RT-PCR with the conserved sequences of AtCBF1 as primers. The coding sequence of AtCBF1 was 642bp ,encoding a polypeptide of 213 amino acids. The nucleotide residue of A in the 252 of ApCBF1 is replaced by T in the same position of AtCBFI. The amino acids sequence of ApCBFI was 100% identical with that in AtCBF1. A recombination vector of pXQ2300-35s ：ApCBF1 was constructed and transferred into Arabidopsis pumila by floral dipping method. Through RT-PCR ,ApCBF1 gene was comfirmed to be overexpressed in different trans-genic plant of T0 generation.